Siglec-15 antibodies in treating bone loss-related disease

ABSTRACT

Novel antibodies and antigen binding fragments that specifically binds to Siglec-15 are described herein In some embodiments, the antibodies or antigen binding fragments may block the biological activity of Siglec-15 and are useful in composition for the treatment of bone loss, more particularly in bone diseases that have increased cell surface expression of Siglec-15, such as conditions where there is an increase in the bone degradative activity of osteoclasts The invention also relates to cells expressing the antibodies or antigen binding fragments such as monoclonal, humanized or chimeric antibodies Additionally, methods of detecting and treating bone loss, bone-related diseases or cancer using the antibodies and fragments are also disclosed.

FIELD OF THE INVENTION

The present invention relates to monoclonal antibodies and antigenbinding fragments thereof that specifically bind to Siglec-15 and theiruse for treating certain diseases including diagnosing, preventing andtreating cancer or bone loss, such as severe or excessive bone lossassociated with bone-related diseases or associated with an increase inosteoclast differentiation or activity. The present invention alsorelates to the use of these antibodies for diagnosis, prevention andtreatment of various other types of diseases where the activity ofosteoclasts is increased.

BACKGROUND OF THE INVENTION

Bone is a dynamic connective tissue comprised of functionally distinctcell populations required to support the structural, mechanical andbiochemical integrity of bone and the human body's mineral homeostasis.The principal cell types involved include, osteoblasts responsible forbone formation and maintaining bone mass, and osteoclasts responsiblefor bone resorption. Osteoblasts and osteoclasts function in a dynamicprocess termed bone remodelling. The development and proliferation ofthese cells from their progenitors is governed by networks of growthfactors and cytokines produced in the bone microenvironment as well asby systemic hormones. Bone remodelling is ongoing throughout thelifetime of the individual and is necessary for the maintenance ofhealthy bone tissue and mineral homeostasis. The process remains largelyin equilibrium and is governed by a complex interplay of systemichormones, peptides and downstream signalling pathway proteins, localtranscription factors, cytokines, growth factors and matrix remodellinggenes.

Any interference or imbalance arising in the bone remodelling processcan produce skeletal disease, with the most common skeletal disorderscharacterized by a net decrease in bone mass. A primary cause of thisreduction in bone mass is an increase in osteoclast number and/oractivity. The most common of such disease, and perhaps the best known,is osteoporosis occurring particularly in women after the onset ofmenopause. In fact osteoporosis is the most significant underlying causeof skeletal fractures in late middle-aged and elderly women. Whileestrogen deficiency has been strongly implicated as a factor inpostmenopausal osteoporosis, there is longstanding evidence thatremodelling is a locally controlled process being that it takes place indiscrete packets throughout the skeleton as first described by Frostover forty years ago (Frost H. M. 1964).

Since bone remodelling takes place in discrete packets, locally producedhormones and enzymes may be more important than systemic hormones forthe initiation of bone resorption and the normal remodelling process.Such local control is mediated by osteoblasts and osteoclasts in themicroenvironment in which they operate. For example, osteoclasts attachto the bone matrix and form a separate compartment between themselvesand the bone surface delimited by a sealing zone formed by a ring ofactin surrounding the ruffled border. Multiple small vesicles transportenzymes toward the bone matrix and internalize partially digested bonematrix. The microenvironment within the sealing zone is rich with thepresence of lysosomal enzymes and is highly acidic compared to thenormal physiological pH of the body. The ruffled border membrane alsoexpresses RANK, the receptor for RANKL, and macrophage-colonystimulating factor (M-CSF) receptor, both of which are responsible forosteoclast differentiation, as well as the calcitonin receptor capableof rapidly inactivating the osteoclast (Baron, R. 2003).

In a complex pattern of inhibition and stimulation, growth hormone,insulin-like growth factor-1, the sex steroids, thyroid hormone,calciotrophic hormones such as PTH and prostaglandin E2, variouscytokines, such as interleukin-1 beta, interleukin-6, and tumor necrosisfactor-alpha, and 1,25-dihydroxyvitamin D (calcitriol) act co-ordinatelyin the bone remodelling process (Jilka et al. 1992; Poli et al. 1994;Srivastava et al. 1998; de Vemejoul 1996).

Thus, it stands to reason that the unique local environments created bythese specialized cells is due to the expression of either uniquegenetic sequences not expressed in other tissues and/or splice variantsof polynucleotides and polypeptides expressed in other tissues. Theisolation and identification of polynucleotides, polypeptides and theirvariants and derivatives specific to osteoclast activity will permit aclearer understanding of the remodelling process and offer tissuespecific therapeutic targets for the treatment of disease states relatedto bone remodelling.

Many diseases linked to bone remodelling are poorly understood,generally untreatable or treatable only to a limited extent. Forexample, osteoarthritis is difficult to treat as there is no cure andtreatment focuses on relieving pain and preventing the affected jointfrom becoming deformed. Non-steroidal anti-inflammatory drugs (NSAIDs)are generally used to relieve pain.

Another example is osteoporosis where the only current medicationsapproved by the FDA for use in the United States are the anti-resorptiveagents that prevent bone breakdown. Estrogen replacement therapy is oneexample of an anti-resorptive agent. Others include alendronate(Fosamax—a biphosphonate anti-resorptive), risedronate (Actonel—abisphosphonate anti-resorptive), raloxifene (Evista—selective estrogenreceptor modulator (SERM)), calcitonin (Calcimar—a hormone), andparathyroid hormone/teriparatide (Forteo—a synthetic version of thehuman hormone, parathyroid hormone, which helps to regulate calciummetabolism).

Bisphosphonates such as alendronate and risedronate bind permanently tothe surface of bone and interfere with osteoclast activity. This allowsthe osteoblasts to outpace the rate of resorption. The most common sideeffects are nausea, abdominal pain and loose bowel movements. However,alendronate is reported to also cause irritation and inflammation of theesophagus, and in some cases, ulcers of the esophagus. Risedronate ischemically different from alendronate and has less likelihood of causingesophagus irritation. However, certain foods, calcium, iron supplements,vitamins and minerals, or antacids containing calcium, magnesium, oraluminum can reduce the absorption of risedronate, thereby resulting inloss of effectiveness.

The most common side effect of Raloxifen and other SERMS (such asTamoxifen) are hot flashes. However, Raloxifene and other hormonereplacement therapies have been shown to increase the risk of bloodclots, including deep vein thrombosis and pulmonary embolism,cardiovascular disease and cancer.

Calcitonin is not as effective in increasing bone density andstrengthening bone as estrogen and the other anti-resorptive agents.Common side effects of either injected or nasal spray calcitonin arenausea and flushing. Patients can develop nasal irritations, a runnynose, or nosebleeds. Injectable calcitonin can cause local skin rednessat the site of injection, skin rash, and flushing.

A situation demonstrative of the link between several disorders ordisease states involving bone remodelling is that of the use ofetidronate (Didronel) first approved by the FDA to treat Paget'sdisease. Paget's disease is a bone disease characterized by a disorderlyand accelerated remodelling of the bone, leading to bone weakness andpain. Didronel has been used ‘off-label’ and in some studies shown toincrease bone density in postmenopausal women with establishedosteoporosis. It has also been found effective in preventing bone lossin patients requiring long-term steroid medications (such as Prednisoneor Cortisone). However, high dose or continuous use of Didronel cancause another bone disease called osteomalacia. Like osteoporosis,osteomalacia can lead to weak bones with increased risk of fractures.Because of osteomalacia concerns and lack of enough studies yetregarding reduction in the rate of bone fractures, the United States FDAhas not approved Didronel for the treatment of osteoporosis.

Osteoporosis therapy has been largely focused on antiresorptive drugsthat reduce the rate of bone loss but emerging therapies show promise inincreasing bone mineral density instead of merely maintaining it orslowing its deterioration. The osteoporosis early stage pipelineconsists largely of drug candidates in new therapeutic classes, inparticular cathepsin K inhibitors, osteoprotegerin and calcilytics aswell as novel bisphosphonates. Some of these are examples where noveldrugs exploiting genomics programs are being developed based on a deeperunderstanding of bone biology and have the potential to change the faceof treatment of bone disorders in the long term.

The present invention describes the use of antibodies specific forSiglec-15 for the diagnosis, prognosis, and treatment (includingprevention) of cancer or bone loss (e.g., severe or excessive bone lossassociated with bone-related disease or associated with an increase inosteoclast differentiation or activity). In particular, the presentinvention relates to the use of anti-Siglec-15 antibodies for inhibitingthe differentiation of osteoclasts.

Sialic-acid-binding immunoglobulin-like lectins (Siglecs) are members ofthe immunoglobulin (Ig) superfamily that have the ability to interactwith sialic acids (McMillan and Crocker, 2008; Crocker et al., 2007).There are several Siglec family members that all share specificstructural features, in particular, displaying an amino-terminal V-setIg domain that binds to sialic acid and a variable number of C2-set Igdomains. These membrane receptors are generally expressed in highlyspecific manners and many of the family members are expressed inhematopoietic cells (McMillan and Crocker, 2008). These proteins arethought to promote cell-cell interactions, mediate signalling, andregulate immune functions through the recognition of glycans (Crocker etal., 2007). Sialic acids are nine-carbon sugars typically located at theends of complex glycoconjugates on the surface of cells. They can beattached to a wide variety of proteins and lipids (McMillan and Crocker,2008).

Siglec-15 is one of the most recently described Siglec family membersthat have a high homology to Siglec-14 (Angata et al., 2007). Theseauthors reported that it preferentially binds to sialyl Tn structure andthat it interacts with DAP12 and DAP10. The functional significance ofthese interactions is not known but it was proposed that Siglec-15probably harbors an activating function (Angata et al., 2007). Despitethese preliminary insights into a potential role in mammals ofSiglec-15, important advances in the understanding of the biologicalfunction of the protein were contributed when the sequence wasidentified as part of a screen to discover novel regulators ofosteoclast differentiation (Sooknanan et al. 2007). In this patentapplication, it was revealed that attenuation of the Siglec-15transcript by RNA interference in a mouse model of osteoclastogenesisresulted in significant reduction of differentiation of precursors inresponse to RANKL treatment. Similar results were disclosed in humanosteoclasts. Furthermore, the studies presented in this disclosure alsoshowed that the localization of Siglec-15 at the cell membrane wasnecessary for its function in osteoclast differentiation. Furthermore, arecent publication showed that the presence of sialic acid at the end ofsurface glycoconjugates was required for proper osteoclastdifferentiation and were probably important for the fusion of osteoclastprecursor cells (Takahata et al., 2007). This last observation creates adirect functional link between sialic acid binding and the expression ofSiglec-15 in differentiating osteoclasts and strongly suggested thatSiglec-15 plays a role in the early differentiation program ofosteoclast precursors.

Thus, the expression profile of Siglec-15, its strong inducibilityduring osteoclast differentiation, its localization at the surface ofthe membrane, and its structural features all contribute to thefeasibility of targeting this protein at the cell surface withmonoclonal antibodies. The only other example of monoclonalantibody-based therapy that target osteoclasts is denosumab, a humanmonoclonal antibody that is specific for RANKL (Ellis et al. 2008). Thepresent invention relates to the use of anti-Siglec-15 antibodies orantigen binding fragments as blockers of osteoclast differentiation inthe detection or treatment of bone loss, especially in the context ofbone-related diseases or in the context of increased osteoclastdifferentiation or activity. The present invention also relates to theuse of antibodies or antigen binding fragments in the detection ortreatment of cancer.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows the PCR-based expression profiling of the Siglec-15 mRNA inhuman differentiating osteoclast samples from six different donors. Alsodepicted is the expression profiling in RNA samples from 30 human normaltissues. As controls, the Siglec-15 expression pattern was compared to awell-known osteoclast marker, cathepsin K (CATK) and the housekeepinggene glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was included tocontrol for the quantity of RNA in each sample.

FIG. 2 shows the expression of the Siglec-15 mRNA in samples isolatedfrom the NCI-60 panel of cancer cell lines.

FIG. 3 presents a Coomassie-stained polyacrylamide gel containing asample of the purified human recombinant Siglec-15 that was expressed asa Fc fusion protein in 293-6E cells. This preparation was used togenerate the monoclonal antibodies disclosed in this patent.

FIG. 4 shows the results of an Fc-Siglec-15 ELISA of the individualmonoclonal antibodies selected from the 96-well plate from Omniclonallibrary #25 containing anti-Siglec-15 Fabs. The wells indicated by boldnumbers contained the exemplary monoclonals 25A1, 25B4, 25B8, 25C1,25D8, 25E5, 25E6, and 25E9. Also shown is an ELISA on the same plateusing the Fc moiety alone to identify those monoclonals that werespecific for the Fc portion of the Fc-Siglec-15 fusion protein.

FIG. 5 presents a scheme that illustrates the steps involved to convertthe mouse Fabs into IgG2 mouse-human chimeric mAbs.

FIG. 6 shows drawings that compare the binding of the mouseanti-Siglec-15 Fabs with the binding of the corresponding IgG2 chimericmonoclonal antibodies for exemplary antibodies 25B4, 25B8, 25C1, 25D8,25E6, and 25E9. The results indicate that the relative binding of theFab variable regions was maintained when transferred to a full humanIgG2 scaffold.

FIG. 7 shows the inhibition of the differentiation of human osteoclastsupon treatment with increasing concentrations of anti-Siglec-15 IgG2chimeric monoclonal antibodies 25B8, 25E6, and 25E9. After treatment,the osteoclasts were stained for TRAP expression.

FIG. 8 shows the inhibition of the differentiation of mouse osteoclastsupon treatment with increasing concentrations of anti-Siglec-15 IgG2chimeric monoclonal antibodies 25B8, 25E6, and 25D8. After treatment,the osteoclasts were stained for TRAP expression.

FIG. 9 shows the comparative binding of the human and mouse Siglec-15 inthe presence of the exemplary antibody 25C8. The result indicates thatthe binding of the antibodies generated against the human Siglec-15 alsointeract with the mouse Siglec-15.

FIGS. 10A, 10B and 10C is a summary of alignment results obtained forselected CDRL1, CDRL2 and CDRL3 sequences (respectively) using theClustalW2 program; where “*” means that the residues in that column areidentical in all sequences in the alignment, “:” means that conservedsubstitutions have been observed and “.” means that semi-conservedsubstitutions are observed. Consensus CDRs were generated using theClustalW program (Larkin M. A., et al., (2007) ClustalW and ClustalXversion 2. Bioinformatics 2007 23(21): 2947-2948).

FIGS. 11A, 11B and 11C is a summary of alignment results obtained forselected CDRH1, CDRH2 and CDRH3 sequences (respectively) using theClustalW2 program; where “*” means that the residues in that column areidentical in all sequences in the alignment, “:” means that conservedsubstitutions have been observed and “.” means that semi-conservedsubstitutions are observed. Consensus CDRs were generated using theClustalW program (Larkin M. A., et al., (2007) ClustalW and ClustalXversion 2. Bioinformatics 2007 23(21): 2947-2948).

FIG. 12 illustrates the ability of the 25E9 candidate antibody that isspecific for Siglec-15 to inhibit the bone resorbing activity ofosteoclasts.

FIGS. 13A, 13B, 13C, 13D and 13E demonstrate that the Siglec-15antibodies can detect the protein by immunoblotting of lysates preparedfrom cells overexpressing the Siglec-15 cDNA (13A), in human (13B) andmouse (13C) osteoclasts, and in U87 glioblastoma cells, and by flowcytometry of intact U87 cells.

FIGS. 14A and 14B shows that the antibodies generated against Siglec-15do not bind other related Siglecs including Siglec-2 and CD33.

FIG. 15 shows an ELISA that demonstrates that the anti-Siglec-15antibodies can inhibit the interaction between Siglec-15 and sialicacids.

SUMMARY OF THE INVENTION

This invention relates to antibodies and antigen binding fragments aswell as kits useful for the treatment (including prevention), detectionand diagnosis of bone loss or cancer. The antibodies and antigen bindingfragments may more particularly be useful for detection ofdifferentiated osteoclast, ovarian cancer cells, renal cancer cells,cancer cells of the central nervous system, prostate cancer cells,melanoma cells, breast cancer cells, lung cancer cells or colon cancercells and diagnosis of bone loss, ovarian cancer, renal cancer, cancerof the central nervous system, prostate cancer, melanoma, breast cancer,lung cancer or colon cancer. The antibodies or antigen binding fragmentof the present invention may also be useful for treating bone loss,ovarian cancer, renal cancer, cancer of the central nervous system,prostate cancer, melanoma, breast cancer, lung cancer or colon cancer.

The antibodies or antigen-binding fragment of the present invention maybind to amino acids 20 to 259 of Siglec-15 (SEQ ID NO.:2) or to acorresponding region of Siglec-15 variant (e.g., SEQ ID NO.:4). Moreparticularly the antibodies or antigen-binding fragment of the presentinvention may bind to amino acids 49 to 165 of Siglec-15 (SEQ ID NO.:2)or to a corresponding region of a Siglec-15 variant (e.g., SEQ IDNO.:4).

The present invention more particularly relates to an isolated antibodyor antigen binding fragment capable of binding to a polypeptide able topromote osteoclast differentiation and of inhibiting an osteoclastdifferentiation activity of the polypeptide.

The antibodies or antigen binding fragments of the present inventionencompass those which bind to amino acids 20 to 259 of SEQ ID NO.:2 orto a variant having at least 80% sequence identity with amino acids 20to 259 of SEQ ID NO.:2.

More particularly, the antibody or antigen binding fragment of thepresent invention may more particularly bind to amino acids 49 to 165 ofSEQ ID NO.:2 or to a variant having at least 80% sequence identity withamino acids 49 to 165 of SEQ ID NO.:2.

More specifically, antibody or antigen binding fragment of the presentinvention may more particularly bind to a polypeptide having at least80% sequence identity with SEQ ID NO.:2.

In accordance with the present invention, the antibody or antigenbinding fragment may therefore interfere with the ability of thepolypeptide to promote osteoclast differentiation or to promote tumorgrowth.

An antibody or antigen binding fragment capable of binding to theextracellular region of SEQ ID NO.:2 or the SEQ ID NO.:2 variant is morespecifically contemplated.

The present invention therefore provides an isolated antibody or antigenbinding fragment capable of binding to a polypeptide able to promoteosteoclast differentiation and having at least 80% sequence identitywith sEQ ID NO.:2 or with amino acids 20 to 259 of SEQ ID NO.:2 (or atleast 80% identity with amino acids 49-165 of SEQ ID NO.:2) ofSialic-acid-binding immunoglobulin-like lectin 15 (Siglec-15; SEQ IDNO.:2), wherein said antibody or antigen binging fragment is capable ofinhibiting osteoclast differentiation, bone resorption (degradation) oris capable of blocking Siglec-15 from binding to a sialic acid.

The antibody or antigen binding fragment of the present invention may becapable of interfering with (inhibiting) differentiation of anosteoclast precursor cell into a differentiated osteoclast.

In accordance with the present invention, the isolated antibody orantigen binding fragment may be, for example, a polyclonal antibody, amonoclonal antibody, a chimeric antibody, a human antibody or a fragmentthereof.

In an exemplary embodiment, the isolated antibody or antigen bindingfragment may be chimeric antibody or a human antibody which may compriseamino acids of a constant region of a human antibody or a fragmentthereof.

The constant region or fragment thereof may be from an IgG1, IgG2, IgG3,or IgG4. In a more specific embodiment, the constant region may be froman IgG2.

Antigen binding fragments which may be particularly be useful include,for example, a FV (scFv), a Fab, a Fab′ or a (Fab′)₂.

The antibody or antigen binding fragment may be produced in or from anisolated mammalian cell (other than an hybridoma cell) or in anhybridoma cell. An exemplary embodiment of an isolated mammalian cell isa human cell.

Production of a monoclonal antibody, a chimeric antibody, a humanantibody or a fragment thereof in an isolated mammalian cell (e.g.,human cell) is particularly contemplated. The chimeric antibody or ahuman antibody thus produced may comprise amino acids of a constantregion of a human antibody or a fragment thereof, including, forexample, a constant region or fragment thereof from an IgG1, IgG2, IgG3,or IgG4. In a more specific embodiment, the constant region may be froman IgG2.

In an aspect of the invention, the antibody or antigen binding fragmentof the present invention may interfere (inhibit) with thedifferentiation of a human osteoclast precursor cell into adifferentiated human osteoclast.

In an exemplary embodiment, the antibody or antigen binding fragment ofthe present invention may interfere (inhibit) with the differentiationof a primary human osteoclast precursor cell into a differentiated humanosteoclast.

Antibodies or antigen binding fragments having such activity mayinclude, for example, a polyclonal antibody, a monoclonal antibody, achimeric antibody, a human antibody or a fragment thereof.

In a more specific embodiment, antibodies or antigen binding fragmentsthat may be capable of having such activity include, for example, amonoclonal antibody, a chimeric antibody, a human antibody or a fragmentthereof.

In an even more specific embodiment, antibodies or antigen bindingfragments that may be capable of having such activity include, forexample, a chimeric antibody, a human antibody or a fragment thereofthat may comprise amino acids of a constant region of a human antibodyor a fragment thereof.

The constant region or fragment thereof of the chimeric or humanantibody may be from an IgG1, IgG2, IgG3, or IgG4. More particularly,the constant region may be from an IgG2.

The antibodies and antigen binding fragments of the present inventionmay also be used to generally target cells expressing or overexpressingSiglec-15, including bone cells and breast, colon, lung, ovarian,prostate, and renal cancer cells as well as melanoma cells and cancercells of the central nervous system.

More particularly, the antibodies and antigen binding fragments may beused to target osteoclasts cells undergoing differentiation.

The present invention provides in one aspect thereof, an isolated orsubstantially purified antibody or antigen binding fragment which may becapable of specific binding to SEQ ID NO:2.

More specifically and in accordance with an embodiment of the invention,the antibody or antigen binding fragment may bind to a domain locatedbetween amino acid 20 and amino acid 259 of SEQ ID NO:2.

In accordance with another embodiment of the invention, the antibody orantigen binding fragment may be capable of binding to an epitopecomprised within amino acid 20 and amino acid 259 of SEQ ID NO:2.

As such, the present invention encompasses diagnostic and/or therapeuticantibodies or antigen binding fragments having specificity for SEQ IDNO:2. Also encompassed by the present invention are antibodies orantigen binding fragments having the same epitope specificity as theantibody of the present invention. A candidate antibody may beidentified by determining whether it will bind to the epitope to whichthe antibodies described herein binds and/or by performing competitionassays with antibodies or antigen binding fragments known to bind to theepitope.

Therefore, another aspect the present invention provides an isolatedantibody or antigen binding fragment capable of competing with theantibody or antigen binding fragment described herein.

In further aspects, the present invention provides method of treatmentand method of detection using the antibody or antigen binding fragmentof the present invention.

The term “antibody” refers to intact antibody, monoclonal or polyclonalantibodies. The term “antibody” also encompasses, multispecificantibodies such as bispecific antibodies. Human antibodies are usuallymade of two light chains and two heavy chains each comprising variableregions and constant regions. The light chain variable region comprises3 CDRs, identified herein as CDRL1, CDRL2 and CDRL3 flanked by frameworkregions. The heavy chain variable region comprises 3 CDRs, identifiedherein as CDRH1, CDRH2 and CDRH3 flanked by framework regions.

The term “antigen-binding fragment”, as used herein, refers to one ormore fragments of an antibody that retain the ability to bind to anantigen (e.g., SEQ ID NO:2 or variants thereof). It has been shown thatthe antigen-binding function of an antibody can be performed byfragments of an intact antibody. Examples of binding fragmentsencompassed within the term “antigen-binding fragment” of an antibodyinclude (i) a Fab fragment, a monovalent fragment consisting of theV_(L), V_(H), C_(L) and C_(H1) domains; (ii) a F(ab′)₂ fragment, abivalent fragment comprising two Fab fragments linked by a disulfidebridge at the hinge region; (iii) a Fd fragment consisting of the V_(H)and C_(H1) domains; (iv) a Fv fragment consisting of the V_(L) and V_(H)domains of a single arm of an antibody, (v) a dAb fragment (Ward et al.,(1989) Nature 341:544-546), which consists of a V_(H) domain; and (vi)an isolated complementarity determining region (CDR), e.g., V_(H) CDR3.Furthermore, although the two domains of the Fv fragment, V_(L) andV_(H), are coded for by separate genes, they can be joined, usingrecombinant methods, by a synthetic linker that enables them to be madeas a single polypeptide chain in which the V_(L) and V_(H) regions pairto form monovalent molecules (known as single chain Fv (scFv); see e.g.,Bird et al. (1988) Science 242:423-426; and Huston et al. (1988) Proc.Natl. Acad. Sci. USA 85:5879-5883). Such single chain antibodies arealso intended to be encompassed within the term “antigen-bindingfragment” of an antibody. Furthermore, the antigen-binding fragmentsinclude binding-domain immunoglobulin fusion proteins comprising (i) abinding domain polypeptide (such as a heavy chain variable region, alight chain variable region, or a heavy chain variable region fused to alight chain variable region via a linker peptide) that is fused to animmunoglobulin hinge region polypeptide, (ii) an immunoglobulin heavychain CH2 constant region fused to the hinge region, and (iii) animmunoglobulin heavy chain CH3 constant region fused to the CH2 constantregion. The hinge region may be modified by replacing one or morecysteine residues with serine residues so as to prevent dimerization.Such binding-domain immunoglobulin fusion proteins are further disclosedin US 2003/0118592 and US 2003/0133939. These antibody fragments areobtained using conventional techniques known to those with skill in theart, and the fragments are screened for utility in the same manner asare intact antibodies.

A typical antigen binding site is comprised of the variable regionsformed by the pairing of a light chain immunoglobulin and a heavy chainimmunoglobulin. The structure of the antibody variable regions is veryconsistent and exhibits very similar structures. These variable regionsare typically comprised of relatively homologous framework regions (FR)interspaced with three hypervariable regions termed ComplementarityDetermining Regions (CDRs). The overall binding activity of the antigenbinding fragment is often dictated by the sequence of the CDRs. The FRsoften play a role in the proper positioning and alignment in threedimensions of the CDRs for optimal antigen binding.

Antibodies and/or antigen binding fragments of the present invention mayoriginate, for example, from a mouse, a rat or any other mammal or fromother sources such as through recombinant DNA technologies.

Further scope, applicability and advantages of the present inventionwill become apparent from the non-restrictive detailed description givenhereinafter. It should be understood, however, that this detaileddescription, while indicating exemplary embodiments of the invention, isgiven by way of example only, with reference to the accompanyingdrawings.

DETAILED DESCRIPTION OF THE INVENTION The Expression Profile ofSiglec-15 in Osteoclasts and Normal Tissues

The present invention relates to the use of monoclonal antibodies totarget osteoclasts found in various bone related disease where severebone loss is observed due to increased activity of the osteoclasts. Inorder to direct the antibodies to the osteoclasts, the identification ofosteoclast-specific antigens that are expressed at the cell surface ofthe cells must be carried out. There are several technologies that areavailable to identify cell-specific antigens and the method that wasused to identify Siglec-15 in differentiating osteoclasts that weretreated with RANKL, an innovative discovery platform called SubtractiveTranscription-based Amplification of mRNA (STAR), is described in thepublished patent application No. PCT/CA2007/000210.

Analysis of the human osteoclast STAR libraries yielded many genes thatencode secreted and cell surface proteins. One of these, termed AB-0326,contained an open reading frame that encoded a polypeptide of 328 aminoacids, corresponding to SEQ ID NO:2 that was encoded by a cDNA of 987base pairs with the nucleotide sequence shown in SEQ ID NO:1. A searchof publicly available databases revealed that the AB-0326 nucleotidesequence was identical to that of a human gene called CD33 antigen-like3 (CD33L3). CD33L3 was later found to be a member of the Siglec familyof sialic acid binding proteins and was renamed Siglec-15 based onhomology to other Siglecs (Crocker et al., 2007). Based on thisinformation, the mouse orthologue was isolated and sequenced and foundto be approximately 85% identical to the human sequence at the aminoacid level. SEQ ID NO:3 and SEQ ID NO:4 show the sequences of cDNA andpolypeptide of the murine Siglec-15, respectively. Bioinformaticanalysis predicted a type I membrane-anchored protein that presents itsfunctional domain to the extracellular compartment. As with other Siglecsequences, an amino-terminal signal peptide (located between amino acids1 and 19 of SEQ ID NO:2) targets the protein to the membrane of cellsand the final processed protein is anchored to the membrane via a singletrans-membrane helix located at the carboxy-terminus (located betweenamino acids 261 and 283 of SEQ ID NO:2). The V-set Ig domain is locatedbetween amino acids 49 and 165 of SEQ ID NO:2 whereas the C2-set Igdomain is located between amino acids 178 and 244 of SEQ ID NO:2.

The present invention relates to the function of Siglec-15 during thedifferentiation of osteoclasts. Previous findings (Sooknanan et al.2007) established that the transcript encoding human Siglec-15 wassignificantly upregulated in response to RANKL. This determination wasperformed on RNA macroarrays that contained spotted total RNA samplesfrom several different human osteoclast differentiation experiments fromdifferent human PBMNC donors. Furthermore, these studies (Sooknanan etal. 2007) revealed that the Siglec-15 transcript was expressed in onlyone normal tissue among a vast panel of 30 human normal tissuesindicating a very high osteoclast specificity of the Siglec-15 geneexpression. Using more sensitive methods such as semi-quantitativeRT-PCR, the expression of the Siglec-15 mRNA was stimulated within oneday of RANKL treatment in many osteoclast samples indicating that thegene was expressed early in osteoclast precursor cells, prior to thecommencement of cell fusion. Finally, the tissue expression profile ofSiglec-15 was assessed by semi-quantitative RT-PCR and found to only beexpressed in a single normal human tissue thus validating the macroarrayresults of Sooknanan et al. Taken together, these expression resultsunderscore the strength of the Applicant's discovery approach in itsability to identify targets, as exemplified by Siglec-15, that arehighly restricted to differentiating osteoclasts.

Based on the expression of Siglec-15 in the early stages ofdifferentiation of osteoclasts, its limited expression in normaltissues, and a critical biological role for Siglec-15 in the activity ofosteoclasts, Siglec-15 was chosen as a therapeutic target for thedevelopment of monoclonal antibodies for the detection, prevention, andtreatment of bone-related diseases such as cancer-induced bone loss andosteoporosis.

Therefore, a variety of anti-Siglec-15 antibodies and immunologicallyfunctional fragments thereof, such as chimeric and humanized monoclonalantibodies, antibody fragments, single chain antibodies, domainantibodies, and polypeptides with an antigen-binding region, fortargeting Siglec-15 are provided.

SEQ ID NO:2 as Antigen and Epitopes Derived from SEQ ID NO:2

In international application No. PCT/CA2007/000210, the Applicant hascome to the unexpected discovery that SEQ ID NO:2 is involved inosteoclast differentiation. This antigen may thus be useful fortargeting cells expressing the antigen in vitro or in vivo and in thedevelopment of detection assays for measuring the antigen in vitro or invivo.

The present invention therefore provides an antigen useful forgenerating specific antibodies and/or specific for cells expressing SEQID NO:2. The antigen or epitope may comprise a fragment of at least 10amino acids (and up to the total length) of SEQ ID NO:2 or of a SEQ IDNO: 2 variant.

An exemplary antigen is the whole SEQ ID NO:2 protein or a variant formhaving at least 80% sequence identity with SEQ ID NO:2 or a fragmentcomprising at least 10 amino acids of SEQ ID NO:2 or of a SEQ ID NO:2variant.

The antigen or the epitope described herein may be fused with a carriersuch as keyhole limpet (KHL), bovine serum albumin (BSA), ovalbumin(OVA) or else in order to generate antibodies and antigen bindingfragments.

The present invention also provides an epitope comprised within aminoacid 20 to 259 of SEQ ID NO:2 to generate antibodies and antigen bindingfragments described herein. The epitope may comprise a fragment of atleast 10 amino acids comprised within amino acids 20 to 259 of SEQ IDNO:2 or a corresponding portion of a SEQ ID NO.:2 variant.

The present invention further provides a composition for generatingantibodies to SEQ ID NO:2 or to a SEQ ID NO:2 variant, the compositionmay comprise an epitope of SEQ ID NO:2 comprised within amino acids 20to 259 of SEQ ID NO:2 or a corresponding portion of a SEQ ID NO:2variant and a carrier.

Exemplary embodiments of compositions are pharmaceutical composition forgenerating antibodies against SEQ ID NO:2 or against a SEQ ID NO:2variant. The pharmaceutical composition may comprise an epitope of SEQID NO:2 comprised within amino acids 20 to 259 of SEQ ID NO:2 or acorresponding portion of a SEQ ID NO:2 variant and a pharmaceuticallyacceptable carrier.

In yet a further aspect the invention provides a method for generatingantibodies against SEQ ID NO:2 or against a SEQ ID NO:2 variant. Themethod may comprise administering a polypeptide comprising an epitope ofSEQ ID NO:2 comprised within amino acids 20 to 259 of SEQ ID NO:2 or acorresponding portion of a SEQ ID NO:2 variant.

In an additional aspect, the present invention provides the use of anepitope of SEQ ID NO:2 comprised within amino acids 20 to 259 of SEQ IDNO:2 or a corresponding portion of a SEQ ID NO:2 variant for generatingantibodies against SEQ ID NO:2 or against a SEQ ID NO:2 variant.

Exemplary embodiments of SEQ ID NO.:2 variant having 80% identity withSEQ ID NO.:2 include for example and without limitation, SEQ ID NO.:4 aswell as other analogues that are published in databases under gene bankaccession numbers or NCBI reference sequence: AAY40743.1,XP_(—)512109.2, XP_(—)001089000.1, XP_(—)601064.4, NP_(—)001094508.1,XP_(—)855238.1, XP_(—)574176.2 and EAX01462.1.

Antibodies and Antigen Binding Fragments that Binds to SEQ ID NO:2 or toSEQ ID NO:2 Variant

Antibodies were initially isolated from Fab libraries for theirspecificity towards the antigen of interest. Comparison of the aminoacid sequences of the light chain variable domains or the heavy chainvariable domains of antibodies showing the greatest characteristicsallowed us to derive consensus sequences within the CDRs and within thevariable regions. The consensus for CDRs are provided in SEQ IDNos:148-158 and 197-210. The consensus for the variable regions areprovided in SEQ ID Nos.:191-196.

The variable regions described herein may be fused with constant regionsof a desired species thereby allowing recognition of the antibody byeffector cells of the desired species. The constant region mayoriginate, for example, from an IgG1, IgG2, IgG3, or IgG4 subtype.Cloning or synthesizing a constant region in frame with a variableregion is well within the scope of a person of skill in the art and maybe performed, for example, by recombinant DNA technology.

In certain embodiments of the present invention, antibodies that bind toSEQ ID NO:2 may be of the IgG1, IgG2, IgG3, or IgG4 subtype. Morespecific embodiments of the invention relates to an antibody of the IgG1subtype. The antibody may be a humanized antibody of the IgG1 subtypethat is biologically active in mediating antibody-dependent cellularcytotoxicity (ADCC), complement-mediated cytotoxicity (CMC), orassociated with immune complexes. The typical ADCC involves activationof natural killer (NK) cells and is reliant on the recognition ofantibody-coated cells by Fc receptors on the surface of the NK cells.The Fc receptors recognize the Fc domain of antibodies such as ispresent on IgG1, which bind to the surface of a target cell, inparticular a bone cell that expresses an antigen, such as SEQ ID NO:2.Once bound to the Fc receptor of IgG the NK cell releases cytokines andcytotoxic granules that enter the target cell and promote cell death bytriggering apoptosis.

The present invention described a collection of antibodies that bind toSEQ ID NO:2. In certain embodiments, the antibodies may be selected fromthe group consisting of polyclonal antibodies, monoclonal antibodiessuch as chimeric or humanized antibodies, antibody fragments such asantigen binding fragments, single chain antibodies, domain antibodies,and polypeptides with an antigen binding region.

The present invention therefore provides in another aspect thereof, anisolated antibody or antigen binding fragment comprising a light chainvariable domain having;

-   -   a. a CDRL1 sequence selected from the group consisting of SEQ ID        NO:69, SEQ ID NO:75, SEQ ID NO:81, SEQ ID NO:87, SEQ ID NO:93,        SEQ ID NO:99, SEQ ID NO:105, SEQ ID NO:111, SEQ ID NO:173, SEQ        ID NO:179 and SEQ ID NO:185;    -   b. a CDRL2 sequence selected from the group consisting of SEQ ID        NO:70, SEQ ID NO:76. SEQ ID NO:82, SEQ ID NO:88, SEQ ID NO:94,        SEQ ID NO:100, SEQ ID NO:106, SEQ ID NO:112, SEQ ID NO:174, SEQ        ID NO:180 and SEQ ID NO:186 and/or;    -   c. a CDRL3 sequence selected from the group consisting of SEQ ID        NO:71, SEQ ID NO:77, SEQ ID NO:83, SEQ ID NO:89, SEQ ID NO:95,        SEQ ID NO:101, SEQ ID NO:107, SEQ ID NO:113, SEQ ID NO:175, SEQ        ID NO:181 and SEQ ID NO:187.

The isolated antibody or antigen binding fragment may also comprise aheavy chain variable domain having;

-   -   a. a CDRH1 sequence selected from the group consisting of SEQ ID        NO:72, SEQ ID NO:78, SEQ ID NO:84, SEQ ID NO:90, SEQ ID NO:96,        SEQ ID NO:102, SEQ ID NO:108, SEQ ID NO:114, SEQ ID NO:176, SEQ        ID NO:182 and SEQ ID NO:188;    -   b. a CDRH2 sequence selected from the group consisting of SEQ ID        NO:73, SEQ ID NO:79, SEQ ID NO:85, SEQ ID NO:91, SEQ ID NO:97,        SEQ ID NO:103, SEQ ID NO:109, SEQ ID NO:115, SEQ ID NO:177, SEQ        ID NO:183 and SEQ ID NO:189 and/or;    -   c. a CDRH3 sequence selected from the group consisting of SEQ ID        NO:74, SEQ ID NO:80, SEQ ID NO:86, SEQ ID NO:92, SEQ ID NO:98,        SEQ ID NO:104, SEQ ID NO:110, SEQ ID NO:116, SEQ ID NO:178, SEQ        ID NO:184 and SEQ ID NO:190.

In a further aspect, the present invention provides an isolated antibodyor antigen binding fragment which may comprise a light chain variabledomain having;

-   -   a) a CDRL1 which may have at least 80% identity with a CDRL1        sequence selected from the group consisting of SEQ ID NO.:148,        SEQ ID NO.:69, SEQ ID NO.:75 and SEQ ID NO.:105    -   b) a CDRL2 which may have at least 80% identity with a CDRL2        sequence selected from the group consisting of SEQ ID NO.:149,        SEQ ID NO.:150, SEQ ID NO.:76, SEQ ID NO.:82 and SEQ ID NO.:106,        or;    -   c) a CDRL3 which may have at least 80% identity with a CDRL3        sequence selected from the group consisting of SEQ ID NO.:151,        SEQ ID NO.:152, SEQ ID NO.:77, SEQ ID NO.:83, SEQ ID NO.:95, SEQ        ID NO.:107 and SEQ ID NO.:152.

In yet a further aspect, the present invention provides an isolatedantibody or antigen binding fragment, wherein the antibody comprises aheavy chain variable domain having;

-   -   a) a CDRH1 which may have at least 80% identity with a CDRH1        sequence selected from the group consisting of SEQ ID NO.:153,        SEQ ID NO.:154, SEQ ID NO.:84, SEQ ID NO.:96 and SEQ ID NO.:102;    -   b) a CDRH2 which may have at least 80% identity with a CDRH2        sequence selected from the group consisting of SEQ ID NO.:155,        SEQ ID NO.:156, SEQ ID NO.:157, SEQ ID NO.:73, SEQ ID NO.:79,        SEQ ID NO.:85, SEQ ID NO.:97, SEQ ID NO.:103 and SEQ ID NO.:109,        or;    -   c) a CDRH3 which may have at least 80% identity with a CDRH3        sequence selected from the group consisting of SEQ ID NO.:158,        SEQ ID NO.:74, SEQ ID NO.:98, SEQ ID NO.:104, SEQ ID NO.:110 and        SEQ ID NO.:116.

In an exemplary embodiment, the antibody or antigen binding fragment maycomprise any individual CDR or a combination of CDR1, CDR2 and/or CDR3of the light chain variable region. The CDR3 may more particularly beselected. Combination may include for example, CDRL1 and CDRL3; CDRL1and CDRL2; CDRL2 and CDRL3 and; CDRL1, CDRL2 and CDRL3.

In another exemplary embodiment, the antibody or antigen bindingfragment may comprise any individual CDR or a combination of CDR1, CDR2and/or CDR3 of the heavy chain variable region. The CDR3 may moreparticularly be selected. Combination may include for example, CDRH1 andCDRH3; CDRH1 and CDRH2; CDRH2 and CDRH3 and; CDRH1, CDRH2 and CDRH3.

In accordance with the present invention, the antibody or antigenbinding fragment may comprise at least two CDRs of a CDRL1, a CDRL2 or aCDRL3.

Also in accordance with the present invention, the antibody or antigenbinding fragment may comprise one CDRL1, one CDRL2 and one CDRL3.

Further in accordance with the present invention, the antibody orantigen binding fragment may comprise:

-   -   a. At least two CDRs of a CDRL1, CDRL2 or CDRL3 and;    -   b. At least two CDRs of a CDRH1, one CDRH2 or one CDRH3.

The antibody or antigen binding fragment may more preferably compriseone CDRL1, one CDRL2 and one CDRL3.

The antibody or antigen binding fragment may also more preferablycomprise one CDRH1, one CDRH2 and one CDRH3.

In another aspect the present invention provides an isolated antibody orantigen binding fragment comprising a heavy chain variable domainhaving;

-   -   a. a CDRH1 sequence selected from the group consisting of SEQ ID        NO:72, SEQ ID NO:78, SEQ ID NO:84, SEQ ID NO:90, SEQ ID NO:96,        SEQ ID NO:102, SEQ ID NO:108, SEQ ID NO:114, SEQ ID NO:176, SEQ        ID NO:182 and SEQ ID NO:188;

b. a CDRH2 sequence selected from the group consisting of SEQ ID NO:73,SEQ ID NO:79, SEQ ID NO:85, SEQ ID NO:91, SEQ ID NO:97, SEQ ID NO:103,SEQ ID NO:109, SEQ ID NO:115, SEQ ID NO:177, SEQ ID NO:183 and SEQ IDNO:189 and/or;

c. a CDRH3 sequence selected from the group consisting of SEQ ID NO:74,SEQ ID NO:80, SEQ ID NO:86, SEQ ID NO:92, SEQ ID NO:98, SEQ ID NO:104,SEQ ID NO:110, SEQ ID NO:116, SEQ ID NO:178, SEQ ID NO:184 and SEQ IDNO:190.

In accordance with the present invention, the antibody or antigenbinding fragment may comprise one CDRH1, one CDRH2 or one CDRH3.

In accordance with the present invention, the antibody or antigenbinding fragment may also comprise one CDRH1, one CDRH2 and one CDRH3.

When only one of the light chain variable domain or the heavy chainvariable domain is available, an antibody or antigen-binding fragmentmay be reconstituted by screening a library of complementary variabledomains using methods known in the art (Portolano et al. The Journal ofImmunology (1993) 150:880-887, Clarkson et al., Nature (1991)352:624-628).

Also encompassed by the present invention are polypeptides or antibodiescomprising variable chains having at least one conservative amino acidsubstitution in at least one of the CDRs described herein.

Also encompassed by the present invention are polypeptides or antibodiescomprising variable chains having at least one conservative amino acidsubstitution in at least two of the CDRs.

Also encompassed by the present invention are polypeptides or antibodiescomprising variable chains having at least one conservative amino acidsubstitution in the 3 CDRs.

Also encompassed by the present invention are polypeptides or antibodiescomprising variable chains having at least two conservative amino acidsubstitution in at least one of the CDRs.

Also encompassed by the present invention are polypeptides or antibodiescomprising variable chains having at least two conservative amino acidsubstitution in at least two of the CDRs.

Also encompassed by the present invention are polypeptides or antibodiescomprising variable chains having at least two conservative amino acidsubstitution in the 3 CDRs.

In another aspect, the present invention relates to a polypeptide,antibody or antigen binding fragment comprising (on a single polypeptidechain or on separate polypeptide chains) at least onecomplementarity-determining region of a light chain variable domain andat least one complementarity-determining region of a heavy chainvariable domain of one of the antibodies or antigen binding fragmentdescribed herein.

The present invention relates in another aspect thereof to antibodiesthat may comprise (on a single polypeptide chain or on separatepolypeptide chains) all six complementarity-determining region (CDR) ofthe antibody or antigen binding fragment described herein.

The antibodies or antigen binding fragment of the present invention mayfurther comprise additional amino acids flanking the amino and/orcarboxy region of the CDR(s). Those additional amino acids may beidentical to the framework regions of the corresponding antibodiesdescribed herein or may include, for example, conservative amino acidsubstitution.

In accordance with an embodiment of the present invention, the antibodyor antigen binding fragment may comprise a CDRL1 sequence comprising orconsisting of formula:

(SEQ ID NO.: 148) RSX_(1a)X_(2a)SLLHSNGX_(3a)TYLY,

Wherein X_(1a) may be, for example, a neutral hydrophilic amino acid;

Wherein X_(2a) may be, for example, lysine or glutamic acid

wherein X_(3a) may be, for example, an hydrophobic amino acid orasparagine.

In a more specific embodiment, X_(1a) may be, for example, serine.

In a more specific embodiment, X_(2a) may be, for example, lysine.

More particularly X_(3a) may be, for example, isoleucine or valine.

In a more specific embodiment, X_(3a) may be isoleucine.

In accordance with yet another embodiment of the present invention, theantibody or antigen binding fragment may comprise a CDRL1 sequencecomprising or consisting of formula:

(SEQ ID NO.: 197) RASX_(a10)NIX_(b10)X_(c10)YLA

Wherein X_(a10) may be any amino acid or for example G or E;

X_(b10) may be any amino acid or for example Y or H, and;

X_(c10) may be any amino acid or for example S or N.

In accordance with yet another embodiment of the present invention, theantibody or antigen binding fragment may comprise a CDRL1 sequencecomprising or consisting of formula: CDRL1 of formulaRSSX_(1x)SLLHSNGX_(2x)TYLY (SEQ ID NO.:201) wherein X_(1x) and X_(2x)are as defined herein.

In accordance with yet another embodiment of the present invention, theantibody or antigen binding fragment may comprise a CDRL1 sequencecomprising or consisting of formula: CDRL1 of formulaRSX_(a6)KSLLHSNGNTYLY (SEQ ID NO.:202) wherein X_(a6) is as definedherein.

The antibody or antigen binding fragment may also comprise, for example,a CDRL1 sequence selected from those comprising or consisting of SEQ IDNO.:75, SEQ ID NO.:69, SEQ ID NO.:105 and other CDRL1 listed in Table 3or Table 5B.

In accordance with another embodiment, the antibody or antigen bindingfragment may comprise a CDRL2 sequence comprising or consisting offormula:

(SEQ ID NO.: 149) X_(1b)MSNLAS,

-   -   wherein X_(1b) may be, for example, a basic amino acid.    -   More particularly, X_(1b) may be, for example, glutamine or        asparagine.    -   In a more specific embodiment, X_(1b) may be glutamine.

In accordance with yet another embodiment, the antibody or antigenbinding fragment may comprise a CDRL2 sequence comprising or consistingof formula:

(SEQ ID NO.: 150) RX_(1c)SNLX_(2c)S,

wherein X_(1c) may be, for example, methionine or threonine and whereinX_(2c) may be, for example, an hydrophobic amino acid.

More particularly, X_(2c) may be, for example, alanine or valine.

In a more specific embodiment, X_(1c) may be, for example, methionine.

In a more specific embodiment, X_(2c) may be, for example, alanine.

In accordance with yet another embodiment of the present invention, theantibody or antigen binding fragment may comprise a CDRL2 sequencecomprising or consisting of formula:

(SEQ ID NO.: 198) NAKTLX_(a11)X_(b11)

X_(a11) may be any amino acid or for example P or A, and;

X_(b11) may be any amino acid or for example an acidic amino acid suchas E or D.

The antibody or antigen binding fragment may also comprise, for example,a CDRL2 sequence selected from those comprising or consisting of SEQ IDNO.:76, SEQ ID NO.:82, SEQ ID NO.:106 and other CDRL2 listed in Table 3or Table 5B.

In accordance with yet another embodiment, the antibody or antigenbinding fragment may comprise a CDRL3 sequence comprising or consistingof formula:

(SEQ ID NO.: 151) X_(1d)QX_(2d)LEX_(3d)PX_(4d)T

wherein X_(1d) may be, for example, an hydrophobic amino acid;

wherein X_(2d) may be, for example, a basic amino acid;

wherein X_(3d) may be, for example, tyrosine or leucine, and;

wherein X_(4d) may be, for example, an aromatic amino acid.

More particularly, X_(1d) may be, for example, methionine or alanine, Ina more specific embodiment, X_(1d) may be, for example, methionine.

More particularly, X_(2d) may be, for example histidine or asparagine.In a more specific embodiment, X_(2d) may be, for example, histidine.

In a more specific embodiment, X_(3d) may be, for example, tyrosine.

More particularly, X_(4d) may be, for example, tyrosine orphenylalanine. In a more specific embodiment, X_(4d) may be, forexample, tyrosine.

In accordance with an additional embodiment, the antibody or antigenbinding fragment may comprise a CDRL3 sequence comprising or consistingof formula:

(SEQ ID NO.: 152) QQWSSNPX_(1e)T

Wherein X_(1e) is proline or leucine.

In accordance with yet another embodiment of the present invention, theantibody or antigen binding fragment may comprise a CDRL3 sequencecomprising or consisting of formula:

(SEQ ID NO.: 199) QHYGX_(a12)PLT

-   -   X_(a12) may be any amino acid or a hydrophobic amino acid such        as for example A or V.

In accordance with a further embodiment of the present invention, theantibody or antigen binding fragment may comprise a CDRL3 sequencecomprising or consisting of formula: X_(a8)QX_(b8)LEX_(c8)PYT (SEQ IDNO.:203) wherein X_(a8), X_(b8) and X_(c8) are as defined herein.

In accordance with yet a further embodiment of the present invention,the antibody or antigen binding fragment may comprise a CDRL3 sequencecomprising or consisting of formula: QHHYGX_(a4)PLT (SEQ ID NO.:204)wherein X_(a4) is as defined herein.

The antibody or antigen binding fragment may also comprise, for example,a CDRL3 sequence selected from those comprising or consisting of SEQ IDNO.:77, SEQ ID NO.:83, SEQ ID NO.:95, SEQ ID NO.:107, SEQ ID NO.:152 andother CDRL3 listed in Table 3 or Table 5B.

In accordance with an additional embodiment, the antibody or antigenbinding fragment may comprise a CDRH1 sequence comprising or consistingof formula:

(SEQ ID NO.: 153) GYTFX_(1f)X_(2f)YX_(3f)MX_(4f)

wherein X_(1f) may be, for example, threonine or asparagine;

wherein X_(2f) may be, for example, threonine, arginine, serine oraspartic acid;

wherein X_(3f) may be, for example, tryptophan or asparagine, asparticacid or glutamic acid, and;

wherein X_(4f) may be, for example, tyrosine, histidine or asparticacid.

In a more specific embodiment, X_(1f) may be, for example, threonine.

In a more specific embodiment, X_(2f) may be, for example, serine.

In a more specific embodiment, X_(3f) may be, for example, tryptophan.

In a more specific embodiment, X_(4f) may be, for example, histidine.

In accordance with yet an additional embodiment, the antibody or antigenbinding fragment may comprise a CDRH1 sequence comprising or consistingof formula:

(SEQ ID NO.: 154) GYTFTDYX_(5f)MH

Wherein X_(5f) may be, for example, an acidic amino acid.

More particularly, X_(5f) may be, for example, glutamic acid or asparticacid. In a more specific embodiment, X_(5f) may be, for example,aspartic acid.

In accordance with a further embodiment of the present invention, theantibody or antigen binding fragment may comprise a CDRH1 sequencecomprising or consisting of formula: GYTFTX_(1l)YWMH (SEQ ID NO.:205)wherein X_(1l) is as defined herein.

In accordance with yet a further embodiment of the present invention,the antibody or antigen binding fragment may comprise a CDRH1 sequencecomprising or consisting of formula: GYTFTDYX_(1s)MH (SEQ ID NO.:208)wherein X_(1s) is as defined herein.

The antibody or antigen binding fragment may also comprise, for example,a CDRH1 sequence selected from those comprising or consisting of SEQ IDNO.:84, SEQ ID NO.:96, SEQ ID NO.:102 and other CDRH1 listed in Table 3or Table 5A.

In accordance with a further embodiment, the antibody or antigen bindingfragment may comprise a CDRH2 sequence comprising or consisting offormula:

(SEQ ID NO.: 155) LINPX_(1g)NX_(2g)RX_(3g)N

Wherein X_(1g) may be, for example, a neutral hydrophilic amino acid;

Wherein X_(2g) may be, for example, alanine or glycine, and;

Wherein X_(3g) may be, for example, proline or threonine.

More particularly, X_(1g) may be, for example, serine or threonine. In amore specific embodiment, X_(1g) may be, for example, threonine.

In a more specific embodiment, X_(2g) may be, for example, glycine.

In a more specific embodiment, X_(3g) may be, for example, threonine.

In accordance with yet a further embodiment, the antibody or antigenbinding fragment may comprise a CDRH2 sequence comprising or consistingof formula:

(SEQ ID NO.: 156) X_(1h)IDPETGGTA

Wherein X_(1h) may be, for example, alanine or threonine.

In accordance with a more specific embodiment, X_(1h) may be, forexample, threonine.

In accordance with yet a further embodiment, the antibody or antigenbinding fragment may comprise a CDRH2 sequence comprising or consistingof formula:

(SEQ ID NO.: 157) EIX_(1i)PX_(2i)X_(3i)SX_(4i)X_(5i)N

Wherein X_(1i) may be, for example, aspartic acid or asparagine;

Wherein X_(2i) may be, for example, aspartic acid or serine;

Wherein X_(3i) may be, for example, aspartic acid or serine;

Wherein X_(4i) may be, for example, tyrosine or threonine, and;

Wherein X_(5i) may be, for example, threonine or isoleucine.

In accordance with yet another embodiment of the present invention, theantibody or antigen binding fragment may comprise a CDRH2 sequencecomprising or consisting of formula:

(SEQ ID NO.: 200) AX_(a13)YPGNGDSR

X_(a13) may be any amino acid or a hydrophobic amino acid such as I orV.

In accordance with an additional embodiment of the present invention,the antibody or antigen binding fragment may comprise a CDRH2 sequencecomprising or consisting of formula: X_(1t)IDPETGGTA (SEQ ID NO.:206)wherein X_(1t) is as defined herein.

In accordance with yet an additional embodiment of the presentinvention, the antibody or antigen binding fragment may comprise a CDRH2sequence comprising or consisting of formula:LINPX_(1m)NX_(2m)RX_(3m)N(SEQ ID NO.:207) wherein X_(1m), X_(2m) and X_(3m) are as definedherein.

In accordance with a further embodiment of the present invention, theantibody or antigen binding fragment may comprise a CDRH2 sequencecomprising or consisting of formula: X_(1t)IDPETGGTA (SEQ ID NO.:209)wherein X_(1t) is as defined herein.

The antibody or antigen binding fragment may also comprise, for example,a CDRH2 sequence selected from those comprising or consisting of SEQ IDNO.:73, SEQ ID NO.:79, SEQ ID NO.:85, SEQ ID NO.:97, SEQ ID NO.:103 andSEQ ID NO.:109 and other CDRH2 listed in Table 3 or Table 5A.

In accordance with an additional embodiment, the antibody or antigenbinding fragment may comprise a CDRH3 sequence comprising or consistingof formula:

(SEQ ID NO.: 158) TX_(1j)FYYX_(2j)X_(3j)X_(4j)NYDVGFAY

Wherein X_(1j) may be, for example, a neutral hydrophilic amino acid;

Wherein X_(2j) may be, for example, a neutral hydrophilic amino acid;

Wherein X_(3j) may be, for example, tyrosine or histidine, and;

Wherein X_(4j) may be, for example, tyrosine or serine.

More particularly, X_(1j) may be, for example, serine or threonine. In amore specific embodiment, X_(1j) may be, for example, serine.

More particularly, X_(2j) may be, for example, serine or threonine. In amore specific embodiment, X_(2j) may be, for example, threonine.

In a more specific embodiment, X_(3j) may be, for example, tyrosine. Ina more specific embodiment, X_(4j) may be, for example, serine.

In accordance with a further embodiment of the present invention, theantibody or antigen binding fragment may comprise a CDRH3 sequencecomprising or consisting of formula:TX_(1v)FYYX_(2v)X_(3v)X_(4v)NYDVGFAY (SEQ ID NO.:210) wherein X_(1v),X_(2v), X_(3v) and X_(4v) are as defined herein.

The antibody or antigen binding fragment may comprise, for example, aCDRH3 sequence selected from those comprising or consisting of SEQ IDNO.:74, SEQ ID NO.:98, SEQ ID NO.:104, SEQ ID NO.:110, SEQ ID NO.:116and other CDRH3 listed in Table 3 or Table 5A.

The framework region of the heavy and/or light chains described hereinmay be derived from one or more of the framework regions illustratedherein. The antibody or antigen binding fragments may thus comprise oneor more of the CDRs described herein (e.g., selected from the specificCDRs or from consensus CDRs SEQ ID NOs.: 148-158 and 197-210) andframework regions originating from the light or heavy chain variableregions illustrated herein.

In an embodiment of the invention, the antibody or antigen bindingfragment of the present invention may comprise a heavy chain variableregion (or a fragment) having formula:

(SEQ ID NO.: 191)X_(1k)X_(2k)QX_(3k)QQX_(4k)X_(5k)X_(6k)EX_(7k)VX_(8k)PGASVKLSCKASGYTFTX_(1l) YW MHWVKQRPGQGLEWIGLINPX _(1m) NX _(2m) RX _(3m)NYNEX_(1n)FX_(2n)X_(3n)KATLTVDKSSSTAYMX_(4n)LSSLTSEDSAVYYCARGGDGDYFDYWGQGTTLTVSSWherein X_(1k) may be for example Q or E;X_(2k) may be any amino acid or a hydrophobic amino acid such as forexample V or I;X_(3k) may be any amino acid or a hydrophobic amino acid such as forexample V or L;X_(4k) may be any amino acid or for example P or S;X_(5k) may be any amino acid or for example R or G;X_(6k) may be any amino acid or for example A or T;X_(7k) may be any amino acid or a hydrophobic amino acid such as forexample L or I;X_(8k) may be any amino acid or a basic amino acid such as for example Ror K;X_(1l) may be any amino acid or a neutral hydrophilic amino acid such asfor example for example S or T;X_(1m) may be any amino acid or a neutral hydrophilic amino acid such asfor example T or S;X_(2m) may be any amino acid or for example G or A;X_(3m) may be any amino acid or for example P or T;X_(1n) may be any amino acid or a basic amino acid such as for example Kor R;X_(2n) may be any amino acid or a basic amino acid such as for example Nor K;X_(3n) may be any amino acid or for example N or a neutral hydrophilicamino acid such as S or T and;X_(4n) may be any amino acid or a basic amino acid such as for example Qor H.

In another embodiment of the invention, the antibody or antigen bindingfragment of the present invention may comprise a heavy chain variableregion (or a fragment) having formula:

(SEQ ID NO.: 192) X₁₀VX₂₀LQQSGAELARPGASVKFSCKASGYTFTRNWIQWVKQRPGQGLEWIGAX _(a13) YPGNGDSRYTQKFKGKATLTADKSSX_(1q)TAYMQLX_(2q)X_(3q)LX_(4q)SEDSAVYYCARLAGNYAYYFDYWGQGTALTVSSWherein X_(1o) may be for example Q or D;X_(2o) may be any amino acid or a basic amino acid such as for example Kor Q;X_(a13) may be any amino acid or a hydrophobic amino acid such as forexample I or V;X_(1q) may be any amino acid or for example S or N;X_(2q) may be any amino acid or for example S or N;X_(3q) may be any amino acid or for example G or S and;X_(4q) may be any amino acid or for example A or S.

In yet another embodiment of the invention, the antibody or antigenbinding fragment of the present invention may comprise a heavy chainvariable region (or a fragment) having formula:

(SEQ ID NO.: 193) X_(1r)X_(2r)X_(3r)LQQSGX_(4r)ELVRPGASVTLSCKASGYTFTDYX_(1s) MHWVKQT PVHGLEWIGX _(1t)IDPETGGTAYNQKFKGKATLTADX_(1u)SSX_(2u)TAYMELS SLTSEDSAVYYCTX _(1v) FYYX_(2v) X _(3v) X _(4v) NYDVGFAYVVGQGTLVTVSAWherein X_(1r) may be for example E or Q;X_(2r) may be any amino acid or a hydrophobic amino acid such as forexample A or I;X_(3r) may be any amino acid or for example Y or Q;X_(4r) may be any amino acid or a hydrophobic amino acid such as forexample A or V;X_(1s) may be any amino acid or an acidic amino acid such as for exampleD or E;X_(1t) may be any amino acid or for example A or T;X_(1u) may be any amino acid or a basic amino acid such as for example Kor R;X_(2u) may be any amino acid or a neutral hydrophilic amino acid such asfor example S or T;X_(1v) may be any amino acid or a neutral hydrophilic amino acid such asfor example S or T;X_(2v) may be any amino acid or a neutral hydrophilic amino acid such asfor example T or S;X_(3v) may be any amino acid or for example Y or H and;X_(4v) may be any amino acid or for example S or Y.

In an additional embodiment, the antibody or antigen binding fragment ofthe present invention may comprise a light chain variable region (or afragment) having formula:

(SEQ ID NO.: 194) DIVMTX_(1w)AX_(2w)FSNPVX_(3w)LGTX_(4w)ASISCRSSX _(1x)SLLHSNGX _(2x) TYLYWYLQKPGQSPQLLIYQMSNLASGVPDRFSX_(1y)SGSGTX_(2y)FTLRISRVEA EDVGVYYCX _(a8)QX _(b8) LEX _(c8) PYTFGX_(a9)GTKLEIKWherein X_(1w) may be any amino acid or a basic amino acid such as forexample Q or H;X_(2w) may be any amino acid or a hydrophobic amino acid such as forexample V or A;X_(3w) may be any amino acid or for example T or I;X_(4w) may be any amino acid or for example S or P;X_(1x) may be any amino acid or for example E or K;X_(2x) may be any amino acid or a hydrophobic amino acid such as forexample V or I;X_(1y) may be any amino acid or for example S or G;X_(2y) may be any amino acid or for example D or A;X_(a8) may be any amino acid or a hydrophobic amino acid such as forexample M or A;X_(b8) may be any amino acid or a basic amino acid such as for example Nor H;X_(c8) may be any amino acid or for example Y or L, and;X_(a9) may be any amino acid or for example G or S.

In a further embodiment, the antibody or antigen binding fragment of thepresent invention may comprise a light chain variable region (or afragment) having formula:

(SEQ ID NO.: 195) X_(1z)IQMTQSPASLSASVGETVTITCRASX _(a10) NIX _(b10) X_(c10) YLAWYQQK QGKSPQLLVYNAKTLX _(a11) X_(b11)GVX_(a3)X_(b3)RFSGSGSGTQX_(c3)SLKIN X_(d3)LQPEDFGSYX_(e3)CQHHYGX_(a4) PLTFGX_(a5)GTKX_(b5)ELKWherein X_(1z) may be any amino acid or for example D or N;X_(a10) may be any amino acid or for example E or G;X_(b10) may be any amino acid or for example Y or H;X_(c10) may be any amino acid or for example S or N;X_(a1111) may be any amino acid or for example P or A;X_(b11) may be any amino acid or an acidic amino acid such as forexample E or D;X_(a3) may be any amino acid or for example P or S;X_(b3) may be any amino acid or for example V or S;X_(c3) may be any amino acid or an aromatic amino acid such as forexample F or Y;X_(d3) may be any amino acid or for example N or S;X_(e3) may be any amino acid or for example H or Y;X_(a4) may be any amino acid or a hydrophobic amino acid such as forexample A or V;X_(a5) may be any amino acid or for example S or A, and;X_(b5) may be any amino acid or a hydrophobic amino acid such as forexample V or L.In yet a further embodiment, the antibody or antigen binding fragment ofthe present invention may comprise a light chain variable region (or afragment) having formula:

(SEQ ID NO.: 196) DIVMTQAAPSVPVTPGESVSISCRSX _(a6)KSLLHSNGNTYLYWFLQRPGQSPQLLIYRMSNLASGVPDRFSGSGSGTAFTLRX_(a7)SRVEAEDVGVYYCMQH LEYPFTFGGGTKLEIKWherein X_(a6) may be any amino acid or a neutral hydrophilic amino acidsuch as for example S or T, and;X_(a7) may be any amino acid or a hydrophobic amino acid such as forexample I or L.Antibodies that Bind to Siglec-15

In certain embodiments of the present invention, antibodies that bind toSiglec-15 are of the IgG1, IgG2, IgG3, or IgG4 subtype. In the preferredembodiment, the antibody is an antibody of the IgG2 subtype. In thepresent embodiment, the antibody is a humanized antibody of the IgG2subtype that is biologically active in blocking the biological activityof normal Siglec-15 function on the surface of osteoclasts. Suchblockage, for example, could prevent the association of Siglec-15 withits substrates, its ligands, itself, or other proteins on adjacentcells.

The present invention discloses a collection of antibodies that bind toSiglec-15. In certain embodiments, the antibodies consist of monoclonalantibodies and immunologically functional fragments thereof, such aschimeric and humanized monoclonal antibodies, antibody fragments, singlechain antibodies, domain antibodies, and polypeptides with anantigen-binding region.

A typical antigen-binding site is comprised of the variable regionsformed by the pairing of a light chain immunoglobulin and a heavy chainimmunoglobulin. The structure of the antibody variable regions is veryconsistent and exhibits very similar structures. These variable regionsare typically comprised of relatively homologous framework regions (FR)interspaced with three hypervariable regions termed ComplementarityDetermining Regions (CDRs). Although the overall binding activity of theantigen binding fragment is dictated by the sequence of the CDRs, theFRs play a critical role in the proper positioning and alignment inthree dimensions of the CDRs for optimal antigen binding.

Table 1 discloses the sequences of the nucleotides and the amino acidscorresponding to the complete light and heavy chain immunoglobulins ofspecific examples of anti-Siglec-15 antibodies.

TABLE 1 Complete sequences of light and heavy chain immunoglobulins thatbind to Siglec-15 Antibody Chain Nucleotide sequence Amino acid sequencedesignation type (SEQ ID NO:) (SEQ ID NO:) 25A1 Light (L) 5 6 25A1 Heavy(H) 7 8 25B4 Light 9 10 25B4 Heavy 11 12 25B8 Light 13 14 25B8 Heavy 1516 25C1 Light 17 18 25C1 Heavy 19 20 25D8 Light 21 22 25D8 Heavy 23 2425E5 Light 25 26 25E5 Heavy 27 28 25E6 Light 29 30 25E6 Heavy 31 32 25E9Light 33 34 25E9 Heavy 35 36

An antibody that can bind Siglec-15 may comprise any one L chain withany one H chain immunoglobulin that is listed in Table 1. In certainembodiments, the light chain of antibody 25A1 may be combined with theheavy chain of 25A1 or the heavy chain of 25B4 to form a completeantibody with Siglec-15-binding activity. In an exemplary embodiment ofthe present invention, the 25A1 L chain may be combined with the 25A1 Hchain, the 25B4 L chain may be combined with the 25B4 H chain, the 25B8L chain may be combined with the 25B8 H chain, the 25C1 L chain may becombined with the 25C1 H chain, the 2D8 L chain may be combined with the25D8 H chain, the 25E5 L chain may be combined with the 25E5 H chain,the 25E6 L chain may be combined with the 25E6 H chain, or the 25E9 Lchain may be combined with the 25E9 H chain. Additionally, some examplesof antibodies or antigen binding fragment may consist of any combinationof two L chains and any two H chains from the list of antibodies listedin Table 1.

The complete nucleotide sequences of the light and heavy immunoglobulinchains of antibody 25A1 are shown in SEQ ID NOS:5 and 7, respectively,and the corresponding amino acid sequences of the light and heavyimmunoglobulin chains of antibody 25A1 are shown in SEQ ID NOS:6 and 8,respectively. Thus, in an exemplary embodiment, an antibody that bindsto Siglec-15 may comprise the light chain amino acid shown in SEQ IDNO:6 combined with the heavy chain amino acid sequence shown in SEQ IDNO:8. In another embodiment, the antibody may comprise two identical orsubstantially identical 25A1 light chains comprising SEQ ID NO:6 or avariant thereof and two identical or substantially identical 25A1 heavychains comprising SEQ ID NO:8 or a variant thereof.

The complete nucleotide sequences of the light and heavy immunoglobulinchains of antibody 25B4 are shown in SEQ ID NOS:9 and 11, respectively,and the corresponding amino acid sequences of the light and heavyimmunoglobulin chains of antibody 25B4 are shown in SEQ ID NOS:10 and12, respectively. Thus, in an exemplary embodiment, an antibody thatbinds to Siglec-15 may comprise the light chain amino acid shown in SEQID NO:10 combined with the heavy chain amino acid sequence shown in SEQID NO:12. In another embodiment, the antibody may comprise two identicalor substantially identical 25B4 light chains comprising SEQ ID NO:10 ora variant thereof and two identical or substantially identical 25B4heavy chains comprising SEQ ID NO:12 or a variant thereof.

The complete nucleotide sequences of the light and heavy immunoglobulinchains of antibody 25B8 are shown in SEQ ID NOS:13 and 15, respectivelyand the corresponding amino acid sequences of the light and heavyimmunoglobulin chains of antibody 25B8 are shown in SEQ ID NOS:14 and16, respectively. Thus, in an exemplary embodiment, an antibody thatbinds to Siglec-15 may comprise the light chain amino acid shown in SEQID NO:14 combined with the heavy chain amino acid sequence shown in SEQID NO:16. In another embodiment, the antibody may comprise two identicalor substantially identical 25B8 light chains comprising SEQ ID NO:14 ora variant thereof and two identical or substantially identical 25B8heavy chains comprising SEQ ID NO:16 or a variant thereof.

The complete nucleotide sequences of the light and heavy immunoglobulinchains of antibody 25C1 are shown in SEQ ID NOS:17 and 19, respectively,and the corresponding amino acid sequences of the light and heavyimmunoglobulin chains of antibody 25C1 are shown in SEQ ID NOS:18 and20, respectively. Thus, in an exemplary embodiment, an antibody thatbinds to Siglec-15 may comprise the light chain amino acid shown in SEQID NO:18 combined with the heavy chain amino acid sequence shown in SEQID NO:20. In another embodiment, the antibody may comprise two identicalor substantially identical 25C1 light chains comprising SEQ ID NO:18 ora variant thereof and two identical or substantially identical 25C1heavy chains comprising SEQ ID NO:20 or a variant thereof.

The complete nucleotide sequences of the light and heavy immunoglobulinchains of antibody 25D8 are shown in SEQ ID NOS:21 and 23, respectively,and the corresponding amino acid sequences of the light and heavyimmunoglobulin chains of antibody 25D8 are shown in SEQ ID NOS:22 and24, respectively. Thus, in an exemplary embodiment, an antibody thatbinds to Siglec-15 may comprise the light chain amino acid shown in SEQID NO:22 combined with the heavy chain amino acid sequence shown in SEQID NO:24. In another embodiment, the antibody may comprise two identicalor substantially identical 25D8 light chains comprising of SEQ ID NO:22or a variant thereof and two identical or substantially identical 25D8heavy chains comprising SEQ ID NO:24 or a variant thereof.

The complete nucleotide sequences of the light and heavy immunoglobulinchains of antibody 25E5 are shown in SEQ ID NOS:25 and 27, respectively,and the corresponding amino acid sequences of the light and heavyimmunoglobulin chains of antibody 25E5 are shown in SEQ ID NOS:26 and28, respectively. Thus, in an exemplary embodiment, an antibody thatbinds to Siglec-15 may comprise the light chain amino acid shown in SEQID NO:26 combined with the heavy chain amino acid sequence shown in SEQID NO:28. In another embodiment, the antibody may comprise two identicalor substantially identical 25E5 light chains comprising SEQ ID NO:26 ora variant thereof and two identical or substantially identical 25E5heavy chains comprising SEQ ID NO:28 or a variant thereof.

The complete nucleotide sequences of the light and heavy immunoglobulinchains of antibody 25E6 are shown in SEQ ID NOS:29 and 31, respectivelyand the corresponding amino acid sequences of the light and heavyimmunoglobulin chains of antibody 25E6 are shown in SEQ ID NOS:30 and32, respectively. Thus, in an exemplary embodiment, an antibody thatbinds to Siglec-15 may comprise the light chain amino acid shown in SEQID NO:30 combined with the heavy chain amino acid sequence shown in SEQID NO:32. In another embodiment, the antibody may comprise two identicalor substantially identical 25E6 light chains comprising SEQ ID NO:30 ora variant thereof and two identical or substantially identical 25E6heavy chains comprising SEQ ID NO:32 or a variant thereof.

The complete nucleotide sequences of the light and heavy immunoglobulinchains of antibody 25E9 are shown in SEQ ID NOS:33 and 35, respectively,and the corresponding amino acid sequences of the light and heavyimmunoglobulin chains of antibody 25E9 are shown in SEQ ID NOS:34 and36, respectively. Thus, in an exemplary embodiment, an antibody thatbinds to Siglec-15 may comprise the light chain amino acid shown in SEQID NO:34 combined with the heavy chain amino acid sequence shown in SEQID NO:36. In another embodiment, the antibody may comprise two identicalor substantially identical 25E9 light chains comprising SEQ ID NO:34 ora variant thereof and two identical or substantially identical 25E9heavy chains comprising SEQ ID NO:36 or a variant thereof.

Variants of other anti-Siglec-15 antibodies or antigen binding fragmentsformed by the combination of light and/or heavy immunoglobulin chainsmay each independently have at least 80%, 85%, 90%, 95%, 97%, or 99%identity to the amino acid sequences listed in Table 1 are alsoprovided. In certain embodiments, the antibody variants may comprise atleast one light chain and one heavy chain. In other instances, theantibody variants may comprise two identical or substantially identicallight chains and two identical or substantially identical heavy chains.In accordance with the present invention, the region of variation may belocated in the constant region or in the variable region. Also inaccordance with the present invention, the region of variation may belocated in the framework region.

Also encompassed by the present invention are antibodies comprising alight chain comprising one of the variable region of the light chainsequence listed in Table 1 or a variant thereof and a heavy chaincomprising one of the variable region of the heavy chain sequence listedin Table 1 or a variant thereof. The light chain and heavy chain maycomprise a constant domain. Combinations of light chains and heavychains of Table 1 are also encompassed by the present invention.

Antibodies or antigen binding fragments that contain the light chain andheavy chain variable regions are also provided in the present invention.Additionally, certain embodiments include antigen binding fragments,variants, and derivatives of these light and heavy chain variableregions.

-   -   Yet other exemplary embodiments of the invention includes an        isolated antibody or antigen binding fragment capable of        specific binding to SEQ ID NO:2 or to a variant thereof, the        antibody comprising:        -   a. the light chain variable domain defined in SEQ ID NO.:38            and the heavy chain variable domain defined in SEQ ID            NO.:40;        -   b. the light chain variable domain defined in SEQ ID NO.:42            and the heavy chain variable domain defined in SEQ ID            NO.:44;        -   c. the light chain variable domain defined in SEQ ID NO.:46            and the heavy chain variable domain defined in SEQ ID            NO.:48;        -   d. the light chain variable domain defined in SEQ ID NO.:50            and the heavy chain variable domain defined in SEQ ID            NO.:52;        -   e. the light chain variable domain defined in SEQ ID NO.:54            and the heavy chain variable domain defined in SEQ ID            NO.:56,        -   f. the light chain variable domain defined in SEQ ID NO.:58            and the heavy chain variable domain defined in SEQ ID            NO.:60;        -   g. the light chain variable domain defined in SEQ ID NO.:62            and the heavy chain variable domain defined in SEQ ID            NO.:64;        -   h. the light chain variable domain defined in SEQ ID NO.:66            and the heavy chain variable domain defined in SEQ ID            NO.:68;

It is to be understood herein, that the light chain variable region ofthe specific combination provided above may be changed for any otherlight chain variable region (especially those of Table 2). Similarly,the heavy chain variable region of the specific combination providedabove may be changed for any other heavy chain variable region(especially those of Table 2).

Antibodies that contain the light chain and heavy chain variable regionsare also provided in the present invention. Additionally, certainembodiments include antigen binding fragments, variants, and derivativesof these light and heavy chain variable regions. Examples of sequencespresent in these light and heavy chain variable regions are disclosed inTable 2.

TABLE 2 Sequences of light and heavy chain variable regions that bind toSiglec-15 Antibody Chain Nucleotide sequence Amino acid sequencedesignation type (SEQ ID NO:) (SEQ ID NO:) 25A1 Light (L) 37 38 25A1Heavy (H) 39 40 25B4 Light 41 42 25B4 Heavy 43 44 25B8 Light 45 46 25B8Heavy 47 48 25C1 Light 49 50 25C1 Heavy 51 52 25D8 Light 53 54 25D8Heavy 55 56 25E5 Light 57 58 25E5 Heavy 59 60 25E6 Light 61 62 25E6Heavy 63 64 25E9 Light 65 66 25E9 Heavy 67 68 25B02 Light 161 162 25B02Heavy 163 164 25D11 Light 165 166 25D11 Heavy 167 168 25E10 Light 169170 25E10 Heavy 171 172

Therefore, antibodies and antigen binding fragments that bind toSiglec-15 may comprise one light chain variable region and one chainheavy variable region of the same designated antibody or in anycombinations. For example, in an exemplary embodiment, an anti-Siglec-15antibody or fragment may comprise the 25A1 light chain variable region(SEQ ID NO:38) and the 25A1 heavy chain variable region (SEQ ID NO:40).In an alternate embodiment, an anti-Siglec-15 antibody or fragment maycomprise the 25A1 light chain variable region (SEQ ID NO:38) and the25B4 heavy chain variable region (SEQ ID NO:44). In another embodiment,the anti-Siglec-15 antibodies may comprise two identical orsubstantially identical light chain variable regions and two identicalor substantially identical heavy chain regions. In yet anotherembodiment, the anti-Siglec-15 antibodies may comprise two differentlight chain variable regions and two different heavy chain regions.

Variants of other anti-Siglec-15 antibodies formed by the combination oflight and/or heavy chain variable regions that each have at least 70%,75%, 80%, 85%, 90%, 95%, 97%, or 99% identity to the amino acidsequences listed in Table 2, Tables 5A and 5B are also provided. Thoseskilled in the art will also recognize that the anti-Siglec-15 antibodyvariants may include conservative amino acid changes, amino acidsubstitutions, deletions, or additions in the amino acid sequences ofthe light and/or heavy chain variable regions listed in Table 2.

TABLE 3 Sequences of the light and heavy chain CDRs SEQ Antibody ChainID Amino acid designation type CDR NO: sequence 25A1 Light (L) CDR1 69SASSSVSYMY 25A1 Light CDR2 70 RTSNLAS 25A1 Light CDR3 71 QQWSSNPLT 25A1Heavy (H) CDR1 72 GYTFTRYWMD 25A1 Heavy CDR2 73 EIDPSDSYTN 25A1 HeavyCDR3 74 ARSGAYSSDYSYDGFAY 25B4 Light CDR1 75 RSSKSLLHSNGITYLY 25B4 LightCDR2 76 QMSNLAS 25B4 Light CDR3 77 MQHLEYPYT 25B4 Heavy CDR1 78GYTFTSYWMH 25B4 Heavy CDR2 79 LINPTNGRTN 25B4 Heavy CDR3 80 ARGGDGDYFDY25B8 Light CDR1 81 RSTKSLLHSNGNTYLY 25B8 Light CDR2 82 RMSNLAS 25B8Light CDR3 83 MQHLEYPFT 25B8 Heavy CDR1 84 GYTFTDYDMH 25B8 Heavy CDR2 85TIDPETGGTA 25B8 Heavy CDR3 86 TTFYYSHYNYDVGFAY 25C1 Light CDR1 87RSSKSLLHSNGNTYLY 25C1 Light CDR2 88 RMSNLAS 25C1 Light CDR3 89 MQHLEYPFT25C1 Heavy CDR1 90 GYTFTDYEMH 25C1 Heavy CDR2 91 AIDPETGGTA 25C1 HeavyCDR3 92 TSFYYTYYNYDVGFAY 25D8 Light CDR1 93 RSSKSLLHSNGITYLY 25D8 LightCDR2 94 QMSNLAS 25D8 Light CDR3 95 AQNLELPYT 25D8 Heavy CDR1 96GYTFTSYWMH 25D8 Heavy CDR2 97 LINPSNARTN 25D8 Heavy CDR3 98 ARGGDGDYFDY25E5 Light CDR1 99 SASSSVSYMY 25E5 Light CDR2 100 RTSNLVS 25E5 LightCDR3 101 QQWSSNPPT 25E5 Heavy CDR1 102 GFDFSKDWMS 25E5 Heavy CDR2 103EINPDSSTIN 25E5 Heavy CDR3 104 SRLEDYEDWYFDV 25E6 Light CDR1 105KASQSVSNAVA 25E6 Light CDR2 106 YTSNRYT 25E6 Light CDR3 107 QQDYTSPWT25E6 Heavy CDR1 108 GYTFNTYNMY 25E6 Heavy CDR2 109 GIDPSNGDTK 25E6 HeavyCDR3 110 TSHTY 25E9 Light CDR1 111 RSTKSLLHSNGNTYLY 25E9 Light CDR2 112RMSNLAS 25E9 Light CDR3 113 MQHLEYPFT 25E9 Heavy CDR1 114 GYTFTDYDMH25E9 Heavy CDR2 115 TIDPETGGTA 25E9 Heavy CDR3 116 TSFYYTYSNYDVGFAY25B02 Light CDR1 173 RASENIYSYLA 25B02 Light CDR2 174 NAKTLPE 25B02Light CDR3 175 HHYGVPLT 25B02 Heavy CDR1 176 GYTFTRNWIQ 25B02 Heavy CDR2177 AIYPGNGDSR 25B02 Heavy CDR3 178 ARLAGNYAYYFDY 25D11 Light CDR1 179RASGNIHNYLA 25D11 Light CDR2 180 NAKTLPE 25D11 Light CDR3 181 QHHYGVPLT25D11 Heavy CDR1 182 GYTFTRNWIQ 25D11 Heavy CDR2 183 AIYPGNGDSR 25D11Heavy CDR3 184 ARLAGNYAYYFDY 25E10 Light CDR1 185 RASGNIHNYLA 25E10Light CDR2 186 NAKTLAD 25E10 Light CDR3 187 QHHYGAPLT 25E10 Heavy CDR1188 GYTFTRNWIQ 25E10 Heavy CDR2 189 AVYPGNGDSR 25E10 Heavy CDR3 190ARLAGNYAYYFDY

In certain embodiments of the present invention, the anti-Siglec-15antibodies or antigen binding fragments may comprise the CDR sequencesshown in Table 3 or have substantial sequence identity to the CDRsequences of Table 3. In an exemplary embodiment, the 25A1anti-Siglec-15 antibody may comprise a light chain variable regioncontaining CDR1, 2, and 3 that are encoded by SEQ ID NOS:68, 69, and 70,respectively, and/or a heavy chain variable region containing CDR1, 2,and 3 that are encoded by SEQ ID NOS:71, 72, and 73, respectively. Inother embodiments the CDR3 region may be sufficient to provide antigenbinding. As such polypeptides comprising the CDRL3 or the CDRH3 or boththe CDRL3 and the CDRH3 are encompassed by the present invention.

Additionally, the anti-Siglec-15 antibodies or antigen binding fragmentsmay include any combination of the CDRs listed in Table 3. For example,the antibodies or antigen binding fragments may include the light chainCDR3 and the heavy chain CDR3. It is understood that the CDRs that arecontained in the anti-Siglec-15 antibodies or antigen binding fragmentsmay be variant CDRs with 80%, 85%, 90%, or 95% sequence identity to theCDR sequences presented in Table 3. Those skilled in the art will alsorecognize that the variants may include conservative amino acid changes,amino acid substitutions, deletions, or additions in the CDR sequenceslisted in Table 3.

Other exemplary embodiments of the invention include an isolatedantibody or antigen binding fragment capable of specific binding to SEQID NO:2 or to a variant thereof (a variant having at least 80% identitywith amino acids 20 to 259 or with amino acids 49-165 of SEQ ID NO.:2),the antibody comprising:

-   -   a. the 3 CDRs of a light chain variable domain listed in Table        5B and the 3 CDRs of a heavy chain variable listed in Table 5A;    -   b. the 3 CDRs of a light chain variable domain defined in SEQ ID        NO.:194 and the 3 CDRs of a heavy chain variable domain defined        in SEQ ID NO.:191;    -   c. the 3 CDRs of a light chain variable domain defined in SEQ ID        NO.:195 and the 3 CDRs of a heavy chain variable domain defined        in SEQ ID NO.:192;    -   d. the 3 CDRs of a light chain variable domain defined in SEQ ID        NO.:196 and the 3 CDRs of a heavy chain variable domain defined        in SEQ ID NO.:193;    -   e. the 3CDRs of a light chain variable domain defined in SEQ ID        NO.:38 and the 3CDRs of a heavy chain variable domain defined in        SEQ ID NO.:40;    -   f. the 3CDRs of a light chain variable domain defined in SEQ ID        NO.:42 and the 3CDRs of a heavy chain variable domain defined in        SEQ ID NO.:44;    -   g. the 3CDRs of a light chain variable domain defined in SEQ ID        NO.:46 and the 3CDRs of a heavy chain variable domain defined in        SEQ ID NO.:48;    -   h. the 3CDRs of a light chain variable domain defined in SEQ ID        NO.:50 and the 3CDRs of a heavy chain variable domain defined in        SEQ ID NO.:52    -   i. the 3CDRs of a light chain variable domain defined in SEQ ID        NO.:54 and the 3CDRs of a heavy chain variable domain defined in        SEQ ID NO.:56;    -   j. the 3CDRs of a light chain variable domain defined in SEQ ID        NO.:58 and the 3CDRs of a heavy chain variable domain defined in        SEQ ID NO.:60;    -   k. the 3CDRs of a light chain variable domain defined in SEQ ID        NO.:62 and the 3CDRs of a heavy chain variable domain defined in        SEQ ID NO.:64;    -   l. the 3CDRs of a light chain variable domain defined in SEQ ID        NO.:66 and the 3CDRs of a heavy chain variable domain defined in        SEQ ID NO.:68;    -   m. the 3 CDRs of a light chain variable domain defined in SEQ ID        NO.:162 and the 3 CDRs of a heavy chain variable domain defined        in SEQ ID NO.:164,    -   n. the 3 CDRs of a light chain variable domain defined in SEQ ID        NO.:166 and the 3 CDRs of a heavy chain variable domain defined        in SEQ ID NO.:168, or;    -   o. the 3 CDRs of a light chain variable domain defined in SEQ ID        NO.:170 and the 3 CDRs of a heavy chain variable domain defined        in SEQ ID NO.:172.

In an additional aspect, the present invention relates to an isolatedantibody or antigen binding fragment capable of specific binding toSiglec-15 or to a variant thereof (a variant having at least 80%identity with amino acids 20 to 259 or with amino acids 49-165 of SEQ IDNO.:2), the antibody comprising:

-   -   a) a light chain variable domain having at least 70% (75%, 80%,        85%, 90%, 95%, 100%) sequence identity with the sequence listed        in Table 5B and heavy chain variable domain having at least 70%        (75%, 80%, 85%, 90%, 95%, 100%) sequence identity with the        sequence listed in Table 5A;    -   b) a light chain variable domain having at least 70% (75%, 80%,        85%, 90%, 95%, 100%) sequence identity with SEQ ID NO.:194 and a        heavy chain variable domain having at least 70% (75%, 80%, 85%,        90%, 95%, 100%) sequence identity with SEQ ID NO.:191;    -   c) a light chain variable domain having at least 70% (75%, 80%,        85%, 90%, 95%, 100%) sequence identity with SEQ ID NO.:195 and a        heavy chain variable domain having at least 70% (75%, 80%, 85%,        90%, 95%, 100%) sequence identity with SEQ ID NO.:192;    -   d) a light chain variable domain having at least 70% (75%, 80%,        85%, 90%, 95%, 100%) sequence identity with SEQ ID NO.:196 and a        heavy chain variable domain having at least 70% (75%, 80%, 85%,        90%, 95%, 100%) sequence identity with SEQ ID NO.:193;    -   e) a light chain variable domain having at least 70% (75%, 80%,        85%, 90%, 95%, 100%) sequence identity with SEQ ID NO.:38 and a        heavy chain variable domain having at least 70% (75%, 80%, 85%,        90%, 95%, 100%) sequence identity with SEQ ID NO.:40,    -   f) a light chain variable domain having at least 70% (75%, 80%,        85%, 90%, 95%, 100%) sequence identity with SEQ ID NO.:42 and a        heavy chain variable domain having at least 70% (75%, 80%, 85%,        90%, 95%, 100%) sequence identity with SEQ ID NO.:44;    -   g) a light chain variable domain having at least 70% (75%, 80%,        85%, 90%, 95%, 100%) sequence identity with SEQ ID NO.:46 and a        heavy chain variable domain having at least 70% (75%, 80%, 85%,        90%, 95%, 100%) sequence identity with SEQ ID NO.:48;    -   h) a light chain variable domain having at least 70% (75%, 80%,        85%, 90%, 95%, 100%) sequence identity with SEQ ID NO.:50 and a        heavy chain variable domain having at least 70% (75%, 80%, 85%,        90%, 95%, 100%) sequence identity with SEQ ID NO.:52;    -   i) a light chain variable domain having at least 70% (75%, 80%,        85%, 90%, 95%, 100%) sequence identity with SEQ ID NO.:54 and a        heavy chain variable domain having at least 70% (75%, 80%, 85%,        90%, 95%, 100%) sequence identity with SEQ ID NO.:56;    -   j) a light chain variable domain having at least 70% (75%, 80%,        85%, 90%, 95%, 100%) sequence identity with SEQ ID NO.:58 and a        heavy chain variable domain having at least 70% (75%, 80%, 85%,        90%, 95%, 100%) sequence identity with SEQ ID NO.:60;    -   k) a light chain variable domain having at least 70% (75%, 80%,        85%, 90%, 95%, 100%) sequence identity with SEQ ID NO.:62 and a        heavy chain variable domain having at least 70% (75%, 80%, 85%,        90%, 95%, 100%) sequence identity with SEQ ID NO.:64,    -   l) a light chain variable domain having at least 70% (75%, 80%,        85%, 90%, 95%, 100%) sequence identity with SEQ ID NO.:66 and a        heavy chain variable domain having at least 70% (75%, 80%, 85%,        90%, 95%, 100%) sequence identity with SEQ ID NO.:68,    -   m) a light chain variable domain having at least 70% (75%, 80%,        85%, 90%, 95%, 100%) sequence identity with SEQ ID NO.:162 and a        heavy chain variable domain having at least 70% (75%, 80%, 85%,        90%, 95%, 100%) sequence identity with SEQ ID NO.:164,    -   n) a light chain variable domain having at least 70% (75%, 80%,        85%, 90%, 95%, 100%) sequence identity with SEQ ID NO.:166 and a        heavy chain variable domain having at least 70% (75%, 80%, 85%,        90%, 95%, 100%) sequence identity with SEQ ID NO.:168, and;    -   o) a light chain variable domain having at least 70% (75%, 80%,        85%, 90%, 95%, 100%) sequence identity with SEQ ID NO.:170 and a        heavy chain variable domain having at least 70% (75%, 80%, 85%,        90%, 95%, 100%) sequence identity with SEQ ID NO.:172.

Again, the light chain variable region of the specific combinationprovided above may be changed for any other light chain variable regiondescribed herein. Similarly, the heavy chain variable region of thespecific combination provided above may be changed for any other heavychain variable region described herein.

Variant Antibody and Antigen Binding Fragments

The present invention also encompasses variants of the antibodies orantigen binding fragments described herein. Variant antibodies orantigen binding fragments included are those having a variation in theamino acid sequence. For example, variant antibodies or antigen bindingfragments included are those having at least one variant CDR (two,three, four, five, six and up to twelve variant CDRs), a variant lightchain variable domain, a variant heavy chain variable domain, a variantlight chain and/or a variant heavy chain. Variant antibodies or antigenbinding fragments included in the present invention are those having,for example, similar or improved binding affinity in comparison with theoriginal antibody or antigen binding fragment.

As used herein the term “variant” applies to any of the sequencedescribed herein and includes for example, a variant CDR (either CDRL1,CDRL2, CDRL3, CDRH1, CDRH2 and/or CDRH3), a variant light chain variabledomain, a variant heavy chain variable domain, a variant light chain, avariant heavy chain, a variant antibody, a variant antigen bindingfragment and a SEQ ID NO.:2 variant.

Variant antibodies or antigen binding fragments encompassed by thepresent invention are those which may comprise an insertion, a deletionor an amino acid substitution (conservative or non-conservative). Thesevariants may have at least one amino acid residue in its amino acidsequence removed and a different residue inserted in its place.

The sites of greatest interest for substitutional mutagenesis includethe hypervariable regions (CDRs), but modifications in the frameworkregion or even in the constant region are also contemplated.Conservative substitutions may be made by exchanging an amino acid (of aCDR, variable chain, antibody, etc.) from one of the groups listed below(group 1 to 6) for another amino acid of the same group.

Generally, mutations in the CDRs may have a greater impact on theantigen binding activity of the antibody or antigen binding fragmentthan mutations in the framework region. Variant antibody or antigenbinding fragments that are encompassed by the present invention arethose which have a substantially identical antigen binding capacity(including similar, identical, or slightly less) to those presentedherein or have a better antigen binding capacity than those presentedherein.

Other exemplary embodiment of conservative substitutions are shown inTable 1A under the heading of “preferred substitutions”. If suchsubstitutions result in a undesired property, then more substantialchanges, denominated “exemplary substitutions” in Table 1A, or asfurther described below in reference to amino acid classes, may beintroduced and the products screened.

It is known in the art that variants may be generated by substitutionalmutagenesis and retain the biological activity of the polypeptides ofthe present invention. These variants have at least one amino acidresidue in the amino acid sequence removed and a different residueinserted in its place. For example, one site of interest forsubstitutional mutagenesis may include a site in which particularresidues obtained from various species are identical. Examples ofsubstitutions identified as “conservative substitutions” are shown inTable 1A. If such substitutions result in a change not desired, thenother type of substitutions, denominated “exemplary substitutions” inTable 1A, or as further described herein in reference to amino acidclasses, are introduced and the products screened.

Substantial modifications in function or immunological identity areaccomplished by selecting substitutions that differ significantly intheir effect on maintaining (a) the structure of the polypeptidebackbone in the area of the substitution, for example, as a sheet orhelical conformation. (b) the charge or hydrophobicity of the moleculeat the target site, or (c) the bulk of the side chain. Naturallyoccurring residues are divided into groups based on common side chainproperties:

-   -   (group 1) hydrophobic: norleucine, methionine (Met), Alanine        (Ala), Valine (Val), Leucine (Leu), Isoleucine (Ile)    -   (group 2) neutral hydrophilic: Cysteine (Cys), Serine (Ser),        Threonine (Thr)    -   (group 3) acidic: Aspartic acid (Asp), Glutamic acid (Glu)    -   (group 4) basic: Asparagine (Asn), Glutamine (Gin), Histidine (H        is), Lysine (Lys), Arginine (Arg)    -   (group 5) residues that influence chain orientation: Glycine        (Gly), Proline (Pro); and    -   (group 6) aromatic: Tryptophan (Trp), Tyrosine (Tyr),        Phenylalanine (Phe)        Non-conservative substitutions will entail exchanging a member        of one of these classes for another.

TABLE 1A Amino acid substitution Original residue Exemplary substitutionConservative substitution Ala (A) Val, Leu, Ile Val Arg (R) Lys, Gln,Asn Lys Asn (N) Gln, His, Lys, Arg, Asp Gln Asp (D) Glu, Asn Glu Cys (C)Ser, Ala Ser Gln (Q) Asn; Glu Asn Glu (E) Asp, Gln Asp Gly (G) Ala AlaHis (H) Asn, Gln, Lys, Arg, Arg Ile (I) Leu, Val, Met, Ala, Phe, Leunorleucine Leu (L) Norleucine, Ile, Val, Met, Ile Ala, Phe Lys (K) Arg,Gln, Asn Arg Met (M) Leu, Phe, Ile Leu Phe (F) Leu, Val, Ile, Ala, TyrTyr Pro (P) Ala Ala Ser (S) Thr Thr Thr (T) Ser Ser Trp (W) Tyr, Phe TyrTyr (Y) Trp, Phe, Thr, Ser Phe Val (V) Ile, Leu, Met, Phe, Ala, Leunorleucine

Variation in the amino acid sequence of the variant antibody or antigenbinding fragment may include an amino acid addition, deletion,insertion, substitution etc., one or more modification in the backboneor side-chain of one or more amino acid, or an addition of a group oranother molecule to one or more amino acids (side-chains or backbone).

Variant antibody or antigen binding fragment may have substantialsequence similarity and/or sequence identity in its amino acid sequencein comparison with that of the original antibody or antigen bindingfragment amino acid sequence. The degree of similarity between twosequences is based upon the percentage of identities (identical aminoacids) and of conservative substitution.

Generally, the degree of similarity and identity between variable chainshas been determined herein using the Blast2 sequence program (Tatiana A.Tatusova, Thomas L. Madden (1999), “Blast 2 sequences—a new tool forcomparing protein and nucleotide sequences”, FEMS Microbiol Lett.174:247-250) using default settings, i.e., blastp program, BLOSUM62matrix (open gap 11 and extension gap penalty 1; gapx dropoff 50, expect10.0, word size 3) and activated filters.

Percent identity will therefore be indicative of amino acids which areidentical in comparison with the original peptide and which may occupythe same or similar position.

Percent similarity will be indicative of amino acids which are identicaland those which are replaced with conservative amino acid substitutionin comparison with the original peptide at the same or similar position.

Variants (i.e., analogues) of the present invention (including VLvariants, VH variants, CDR variants, antibody variants, polypeptidevariants, etc.) therefore comprise those which may have at least 70%,75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%,93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity with anoriginal sequence or a portion of an original sequence.

In accordance with the present invention, a SEQ ID NO.:2 variantincludes a polypeptide having a region at least 80% identical with aminoacids 49-165 or with amino acids 20 to 259 of SEQ ID NO.:2. Variants ofSEQ ID NO.:2 also include polypeptides having at least 80% sequenceidentity with SEQ ID NO.:2.

Exemplary embodiments of variants are those having at least 81% sequenceidentity to a sequence described herein and 81%, 82%, 83%, 84%, 85%,86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or100% sequence similarity with an original sequence or a portion of anoriginal sequence.

Other exemplary embodiments of variants are those having at least 82%sequence identity to a sequence described herein and 82%, 83%, 84%, 85%,86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or100% sequence similarity with an original sequence or a portion of anoriginal sequence.

Further exemplary embodiments of variants are those having at least 85%sequence identity to a sequence described herein and 85%, 86%, 87%, 88%,89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequencesimilarity with an original sequence or a portion of an originalsequence.

Other exemplary embodiments of variants are those having at least 90%sequence identity to a sequence described herein and 90%, 91%, 92%, 93%,94%, 95%, 96%, 97%, 98%, 99% or 100% sequence similarity with anoriginal sequence or a portion of an original sequence.

Additional exemplary embodiments of variants are those having at least95% sequence identity to a sequence described herein and 95%, 96%, 97%,98%, 99% or 100% sequence similarity with an original sequence or aportion of an original sequence.

Yet additional exemplary embodiments of variants are those having atleast 97% sequence identity to a sequence described herein and 97%, 98%,99% or 100% sequence similarity with an original sequence or a portionof an original sequence.

For a purpose of concision the applicant provides herein a Table 1Billustrating exemplary embodiments of individual variants encompassed bythe present invention and comprising the specified % sequence identityand % sequence similarity. Each “X” is to be construed as defining agiven variant.

TABLE 1B Percent (%) sequence identity 80 81 82 83 84 85 86 87 88 89 9091 92 93 94 95 96 97 98 99 100 Percent (%) 80 X sequence 81 X Xsimilarity 82 X X X 83 X X X X 84 X X X X X 85 X X X X X X 86 X X X X XX X 87 X X X X X X X X 88 X X X X X X X X X 89 X X X X X X X X X X 90 XX X X X X X X X X X 91 X X X X X X X X X X X X 92 X X X X X X X X X X XX X 93 X X X X X X X X X X X X X X 94 X X X X X X X X X X X X X X X 95 XX X X X X X X X X X X X X X X 96 X X X X X X X X X X X X X X X X X 97 XX X X X X X X X X X X X X X X X X 98 X X X X X X X X X X X X X X X X X XX 99 X X X X X X X X X X X X X X X X X X X X 100 X X X X X X X X X X X XX X X X X X X X X

As used herein, the term “identical” means that a sequence share 100%sequence identity with another sequence.

As used herein, the term “substantially identical” means that a sequenceshare 70%, 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%,91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence identity withanother sequence or a portion of another sequence.

The present invention encompasses CDRs, light chain variable domains,heavy chain variable domains, light chains, heavy chains, antibodiesand/or antigen binding fragments which comprise at least 80% identitywith the sequence described herein.

Exemplary embodiments of the antibody or antigen binding fragment of thepresent invention are those comprising a light chain variable domaincomprising a sequence selected from the group consisting of a sequenceat least 70% identical (including 80%, 85%, 90%, 95% and 100% identical)to SEQ ID NO.:38, a sequence at least 70% identical (including 80%, 85%,90%, 95% and 100% identical) to SEQ ID NO.:42, a sequence at least 70%identical (including 80%, 85%, 90%, 95% and 100% identical) to SEQ IDNO.:46, a sequence at least 70% identical (including 80%, 85%, 90%, 95%and 100% identical) to SEQ ID NO.:50, a sequence at least 70% identical(including 80%, 85%, 90%, 95% and 100% identical) to SEQ ID NO.:54, asequence at least 70% identical (including 80%, 85%, 90%, 95% and 100%)identical to SEQ ID NO.:58, a sequence at least 70% identical (including80%, 85%, 90%, 95% and 100% identical) to SEQ ID NO.:62, a sequence 70%identical (including at least 80%, 85%, 90%, 95% and 100% identical) toSEQ ID NO.:66, a sequence 70% identical (including at least 80%, 85%,90%, 95% and 100% identical) to SEQ ID NO.:162, a sequence 70% identical(including at least 80%, 85%, 90%, 95% and 100% identical) to SEQ IDNO.:166 and a sequence 70% identical (including at least 80%, 85%, 90%,95% and 100% identical) to SEQ ID NO.:170.

These light chain variable domain may comprise a CDRL1 sequence at least80% identical to SEQ ID NO:69, a CDRL2 sequence at least 80% identicalto SEQ ID NO: 70 and a CDRL3 sequence at least 80% identical to SEQ IDNO: 71.

In an exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRL1 sequence which may be atleast 90% identical to SEQ ID NO:69.

In another exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRL1 sequence which may be100% identical to SEQ ID NO:69.

In another exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRL2 sequence at least 90%identical to SEQ ID NO: 70.

In yet another exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRL2 sequence which may be100% identical to SEQ ID NO: 70.

In another exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRL3 sequence which may be atleast 90% identical to SEQ ID NO: 71.

In an additional exemplary embodiment of the present invention, any ofthe antibodies provided herein may comprise a CDRL3 sequence which maybe 100% identical to SEQ ID NO: 71.

The light chain variable domain listed above may comprise a CDRL1sequence at least 80% identical to SEQ ID NO: 75, a CDRL2 sequence atleast 80 identical to SEQ ID NO: 76 and a CDRL3 sequence at least 80%identical to SEQ ID NO: 77.

In an exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRL1 sequence which may be atleast 90% identical to SEQ ID NO: 75.

In a further exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRL1 sequence which may be100% identical to SEQ ID NO: 75.

In yet a further exemplary embodiment of the present invention, any ofthe antibodies provided herein may comprise a the CDRL2 sequence whichmay be at least 90% identical to SEQ ID NO: 76.

In another exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRL2 sequence which may be100% identical to SEQ ID NO: 76.

In another exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRL3 sequence which may be atleast 90% identical to SEQ ID NO: 77.

In yet another exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRL3 sequence which may be100% identical to SEQ ID NO: 77.

The light chain variable domain listed above may comprise a CDRL1sequence at least 80% identical to SEQ ID NO: 81, a CDRL2 sequence atleast 80% identical to SEQ ID NO: 82 and a CDRL3 sequence at least 80%identical to SEQ ID NO: 83.

In an exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRL1 sequence which may be atleast 90% identical to SEQ ID NO: 81.

In a further exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRL1 sequence which may be100% identical to SEQ ID NO: 81.

In yet a further exemplary embodiment of the present invention, any ofthe antibodies provided herein may comprise a the CDRL2 sequence whichmay be at least 90% identical to SEQ ID NO: 82.

In another exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRL2 sequence which may be100% identical to SEQ ID NO: 82.

In another exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRL3 sequence which may be atleast 90% identical to SEQ ID NO: 83.

In yet another exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRL3 sequence which may be100% identical to SEQ ID NO: 83.

The light chain variable domain listed above may comprise a CDRL1sequence at least 80% identical to SEQ ID NO: 87, a CDRL2 sequence atleast 80% identical to SEQ ID NO: 88 and a CDRL3 sequence at least 80%identical to SEQ ID NO: 89.

In an exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRL1 sequence which may be atleast 90% identical to SEQ ID NO: 87.

In a further exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRL1 sequence which may be100% identical to SEQ ID NO: 87.

In yet a further exemplary embodiment of the present invention, any ofthe antibodies provided herein may comprise a the CDRL2 sequence whichmay be at least 90% identical to SEQ ID NO: 88.

In another exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRL2 sequence which may be100% identical to SEQ ID NO: 88.

In another exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRL3 sequence which may be atleast 90% identical to SEQ ID NO: 89.

In yet another exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRL3 sequence which may be100% identical to SEQ ID NO: 89.

The light chain variable domain listed above may comprise a CDRL1sequence at least 80% identical to SEQ ID NO: 93, a CDRL2 sequence atleast 80% identical to SEQ ID NO: 94 and a CDRL3 sequence at least 80%identical to SEQ ID NO: 95.

In an exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRL1 sequence which may be atleast 90% identical to SEQ ID NO: 93.

In a further exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRL1 sequence which may be100% identical to SEQ ID NO: 93.

In yet a further exemplary embodiment of the present invention, any ofthe antibodies provided herein may comprise a the CDRL2 sequence whichmay be at least 90% identical to SEQ ID NO: 94.

In another exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRL2 sequence which may be100% identical to SEQ ID NO: 94.

In another exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRL3 sequence which may be atleast 90% identical to SEQ ID NO: 95.

In yet another exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRL3 sequence which may be100% identical to SEQ ID NO: 95.

The light chain variable domain listed above may comprise a CDRL1sequence at least 80% identical to SEQ ID NO: 99, a CDRL2 sequence atleast 80% identical to SEQ ID NO: 100 and a CDRL3 sequence at least 80%identical to SEQ ID NO: 101.

In an exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRL1 sequence which may be atleast 90% identical to SEQ ID NO: 99.

In a further exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRL1 sequence which may be100% identical to SEQ ID NO: 99.

In yet a further exemplary embodiment of the present invention, any ofthe antibodies provided herein may comprise a the CDRL2 sequence whichmay be at least 90% identical to SEQ ID NO: 100.

In another exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRL2 sequence which may be100% identical to SEQ ID NO: 100.

In another exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRL3 sequence which may be atleast 90% identical to SEQ ID NO: 101.

In yet another exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRL3 sequence which may be100% identical to SEQ ID NO: 101.

The light chain variable domain listed above may comprise a CDRL1sequence at least 80% identical to SEQ ID NO: 105, a CDRL2 sequence atleast 80% identical to SEQ ID NO: 106 and a CDRL3 sequence at least 80%identical to SEQ ID NO: 107.

In an exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRL1 sequence which may be atleast 90% identical to SEQ ID NO: 105.

In a further exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRL1 sequence which may be100% identical to SEQ ID NO: 105.

In yet a further exemplary embodiment of the present invention, any ofthe antibodies provided herein may comprise a CDRL2 sequence which maybe at least 90% identical to SEQ ID NO: 106.

In another exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRL2 sequence which may be100% identical to SEQ ID NO: 106.

In another exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRL3 sequence which may be atleast 90% identical to SEQ ID NO: 107.

In yet another exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRL3 sequence which may be100% identical to SEQ ID NO: 107.

The light chain variable domain listed above may comprise a CDRL1sequence at least 80% identical to SEQ ID NO: 111, a CDRL2 sequence atleast 80% identical to SEQ ID NO: 112 and a CDRL3 sequence at least 80%identical to SEQ ID NO: 113.

In an exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRL1 sequence which may be atleast 90% identical to SEQ ID NO: 111.

In a further exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRL1 sequence which may be100% identical to SEQ ID NO: 111.

In yet a further exemplary embodiment of the present invention, any ofthe antibodies provided herein may comprise a CDRL2 sequence which maybe at least 90% identical to SEQ ID NO: 112.

In another exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRL2 sequence which may be100% identical to SEQ ID NO: 112.

In another exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRL3 sequence which may be atleast 90% identical to SEQ ID NO: 113.

In another exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRL3 sequence which may be atleast 100% identical to SEQ ID NO: 113.

The light chain variable domain listed above may comprise a CDRL1sequence at least 80% identical to SEQ ID NO.:173, a CDRL2 sequence atleast 80% identical to SEQ ID NO.:174 and a CDRL3 sequence at least 80%identical to SEQ ID NO.:175.

In an exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRL1 sequence which may be atleast 90% identical to SEQ ID NO: 173.

In a further exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRL1 sequence which may be100% identical to SEQ ID NO: 173.

In yet a further exemplary embodiment of the present invention, any ofthe antibodies provided herein may comprise a CDRL2 sequence which maybe at least 90% identical to SEQ ID NO: 174

In another exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRL2 sequence which may be100% identical to SEQ ID NO: 174.

In another exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRL3 sequence which may be atleast 90% identical to SEQ ID NO: 175.

In another exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRL3 sequence which may be atleast 100% identical to SEQ ID NO: 175.

The light chain variable domain listed above may comprise a CDRL1sequence at least 80% identical to SEQ ID NO.:179, a CDRL2 sequence atleast 80% identical to SEQ ID NO.:180 and a CDRL3 sequence at least 80%identical to SEQ ID NO.:181.

In an exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRL1 sequence which may be atleast 90% identical to SEQ ID NO: 179.

In a further exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRL1 sequence which may be100% identical to SEQ ID NO: 179.

In yet a further exemplary embodiment of the present invention, any ofthe antibodies provided herein may comprise a CDRL2 sequence which maybe at least 90% identical to SEQ ID NO: 180.

In another exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRL2 sequence which may be100% identical to SEQ ID NO: 180.

In another exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRL3 sequence which may be atleast 90% identical to SEQ ID NO: 181.

In another exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRL3 sequence which may be atleast 100% identical to SEQ ID NO: 181.

The light chain variable domain listed above may comprise a CDRL1sequence at least 80% identical to SEQ ID NO.:185, a CDRL2 sequence atleast 80% identical to SEQ ID NO.:186 and a CDRL3 sequence at least 80%identical to SEQ ID NO.:187.

In an exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRL1 sequence which may be atleast 90% identical to SEQ ID NO: 185.

In a further exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRL1 sequence which may be100% identical to SEQ ID NO: 185.

In yet a further exemplary embodiment of the present invention, any ofthe antibodies provided herein may comprise a CDRL2 sequence which maybe at least 90% identical to SEQ ID NO: 186.

In another exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRL2 sequence which may be100% identical to SEQ ID NO: 186.

In another exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRL3 sequence which may be atleast 90% identical to SEQ ID NO: 187.

In another exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRL3 sequence which may be atleast 100% identical to SEQ ID NO: 187. In an exemplary embodiment, theantibody or antigen binding fragment may comprise a heavy chain variabledomain comprising a sequence selected from the group consisting of asequence at least 70% identical (including 80%, 85%, 90%, 95%, 100%identical) to SEQ ID NO:40, a sequence at least 70% identical (including80% identical) to SEQ ID NO:44, a sequence at least 70% identical(including 80%, 85%, 90%, 95%, 100% identical) to SEQ ID NO:48, asequence at least 70% identical (including 80%, 85%, 90%, 95%, 100%identical) to SEQ ID NO:52, a sequence at least 70% identical (including80%, 85%, 90%, 95%, 100% identical) to SEQ ID NO:56, a sequence at least70% identical (including 80%, 85%, 90%, 95%, 100% identical) to SEQ IDNO:60, a sequence at least 70% identical (including 80%, 85%, 90%, 95%,100%) identical to SEQ ID NO:64, a sequence at least 70% identical(including 80%, 85%, 90%, 95%, 100% identical) to SEQ ID NO:68, asequence at least 70% identical (including 80%, 85%, 90%, 95%, 100%identical) to SEQ ID NO:164, a sequence at least 70% identical(including 80%, 85%, 90%, 95%, 100% identical) to SEQ ID NO:168 and asequence at least 70% identical (including 80%, 85%, 90%, 95%, 100%identical) to SEQ ID NO:172.

These heavy chain variable domain may comprise a CDRH1 sequence at least80% identical to SEQ ID NO:72, a CDRH2 sequence at least 80% identicalto SEQ ID NO:73 and a CDRH3 sequence at least 80% identical to SEQ IDNO:74.

In an exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRH1 sequence which may be atleast 90% identical to SEQ ID NO:72.

In another exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRH1 sequence which may be100% identical to SEQ ID NO:72.

In yet another exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRH2 sequence which may be atleast 90% identical to SEQ ID NO:73.

In a further exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRH2 sequence which may be100% identical to SEQ ID NO:73.

In yet a further exemplary embodiment of the present invention, any ofthe antibodies provided herein may comprise a CDRH3 sequence which maybe at least 90% identical to SEQ ID NO:74.

In an additional exemplary embodiment of the present invention, any ofthe antibodies provided herein may comprise a CDRH3 sequence which maybe 100% identical to SEQ ID NO:74.

The heavy chain variable domain listed above may comprise a CDRH1sequence at least 80% identical to SEQ ID NO:78, a CDRH2 sequence atleast 80% identical to SEQ ID NO:79 and a CDRH3 sequence at least 80%identical to SEQ ID NO:80.

In an exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRH1 sequence which may be atleast 90% identical to SEQ ID NO:78.

In another exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRH1 sequence which may be100% identical to SEQ ID NO:78.

In an additional exemplary embodiment of the present invention, any ofthe antibodies provided herein may comprise a CDRH2 sequence which maybe at least 90% identical to SEQ ID NO:79.

In a further exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRH2 sequence which may be100% identical to SEQ ID NO:79.

In another exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRH3 sequence which may be atleast 90% identical to SEQ ID NO:80.

In another exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRH3 sequence which may be100% identical to SEQ ID NO:80.

The light chain variable domain listed above may comprise a CDRH1sequence at least 80% identical to SEQ ID NO: 84, a CDRH2 sequence atleast 80% identical to SEQ ID NO: 85 and a CDRH3 sequence at least 80%identical to SEQ ID NO: 86.

In an exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRH1 sequence which may be atleast 90% identical to SEQ ID NO: 84.

In a further exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRH1 sequence which may be100% identical to SEQ ID NO: 84.

In yet a further exemplary embodiment of the present invention, any ofthe antibodies provided herein may comprise a CDRH2 sequence which maybe at least 90% identical to SEQ ID NO: 85.

In another exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRH2 sequence which may be100% identical to SEQ ID NO: 85.

In another exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRH3 sequence which may be atleast 90% identical to SEQ ID NO: 86.

In yet another exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRH3 sequence which may be100% identical to SEQ ID NO: 86.

The light chain variable domain listed above may comprise a CDRH1sequence at least 80% identical to SEQ ID NO: 90, a CDRH2 sequence atleast 80% identical to SEQ ID NO: 91 and a CDRH3 sequence at least 80%identical to SEQ ID NO: 92.

In an exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRH1 sequence which may be atleast 90% identical to SEQ ID NO: 90.

In a further exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRH1 sequence which may be100% identical to SEQ ID NO: 90.

In yet a further exemplary embodiment of the present invention, any ofthe antibodies provided herein may comprise a CDRH2 sequence which maybe at least 90% identical to SEQ ID NO: 91.

In another exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRH2 sequence which may be100% identical to SEQ ID NO: 91.

In another exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRH3 sequence which may be atleast 90% identical to SEQ ID NO: 92.

In yet another exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRH3 sequence which may be100% identical to SEQ ID NO: 92.

The light chain variable domain listed above may comprise a CDRH1sequence at least 80% identical to SEQ ID NO: 96, a CDRH2 sequence atleast 80% identical to SEQ ID NO: 97 and a CDRH3 sequence at least 80%identical to SEQ ID NO: 98.

In an exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRH1 sequence which may be atleast 90% identical to SEQ ID NO: 96.

In a further exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRH1 sequence which may be100% identical to SEQ ID NO: 96.

In yet a further exemplary embodiment of the present invention, any ofthe antibodies provided herein may comprise a CDRH2 sequence which maybe at least 90% identical to SEQ ID NO: 97.

In another exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRH2 sequence which may be100% identical to SEQ ID NO: 97.

In another exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRH3 sequence which may be atleast 90% identical to SEQ ID NO: 98.

In yet another exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRH3 sequence which may be100% identical to SEQ ID NO: 98.

The light chain variable domain listed above may comprise a CDRH1sequence at least 80% identical to SEQ ID NO: 102, a CDRH2 sequence atleast 80% identical to SEQ ID NO: 103 and a CDRH3 sequence at least 80%identical to SEQ ID NO: 104.

In an exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRH1 sequence which may be atleast 90% identical to SEQ ID NO: 102.

In a further exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRH1 sequence which may be100% identical to SEQ ID NO: 102.

In yet a further exemplary embodiment of the present invention, any ofthe antibodies provided herein may comprise a CDRH2 sequence which maybe at least 90% identical to SEQ ID NO: 103.

In another exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRH2 sequence which may be100% identical to SEQ ID NO: 103.

In another exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRH3 sequence which may be atleast 90% identical to SEQ ID NO: 104.

In yet another exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRH3 sequence which may be100% identical to SEQ ID NO: 104.

These heavy chain variable domain may comprise a CDRH1 sequence at least80% identical to SEQ ID NO:108, a CDRH2 sequence at least 80% identicalto SEQ ID NO:109 and a CDRH3 sequence at least 80% identical to SEQ IDNO:110. In an exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRH1 sequence which may be atleast 90% identical to SEQ ID NO: 108.

In another exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRH1 sequence which may be100% identical to SEQ ID NO: 108.

In yet another exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRH2 sequence which may be atleast 90% identical to SEQ ID NO: 109.

In a further exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRH2 sequence which may be100% identical to SEQ ID NO: 109.

In yet a further exemplary embodiment of the present invention, any ofthe antibodies provided herein may comprise a CDRH3 sequence which maybe at least 90% identical to SEQ ID NO: 110.

In an additional exemplary embodiment of the present invention, any ofthe antibodies provided herein may comprise a CDRH3 sequence which maybe 100% identical to SEQ ID NO: 110.

These heavy chain variable domain may comprise a CDRH1 sequence at least80% identical to SEQ ID NO:114, a CDRH2 sequence at least 80% identicalto SEQ ID NO:115 and a CDRH3 sequence at least 80% identical to SEQ IDNO:116.

In an exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRH1 sequence which may be atleast 90% identical to SEQ ID NO: 114.

In another exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRH1 sequence which may be100% identical to SEQ ID NO: 114.

In yet another exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRH2 sequence which may be atleast 90% identical to SEQ ID NO: 115.

In a further exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRH2 sequence which may be100% identical to SEQ ID NO: 115.

In yet a further exemplary embodiment of the present invention, any ofthe antibodies provided herein may comprise a CDRH3 sequence which maybe at least 90% identical to SEQ ID NO: 116.

In an additional exemplary embodiment of the present invention, any ofthe antibodies provided herein may comprise a CDRH3 sequence which maybe 100% identical to SEQ ID NO: 116.

These heavy chain variable domains may comprise a CDRH1 sequence atleast 80% identical to SEQ ID NO.:176, a CDRH2 sequence at least 80%identical to SEQ ID NO.:177 and a CDRH3 sequence at least 80% identicalto SEQ ID NO.:178.

In an exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRH1 sequence which may be atleast 90% identical to SEQ ID NO.: 176.

In another exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRH1 sequence which may be100% identical to SEQ ID NO.: 176.

In yet another exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRH2 sequence which may be atleast 90% identical to SEQ ID NO.: 177.

In a further exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRH2 sequence which may be100% identical to SEQ ID NO.: 177.

In yet a further exemplary embodiment of the present invention, any ofthe antibodies provided herein may comprise a CDRH3 sequence which maybe at least 90% identical to SEQ ID NO.: 178.

In an additional exemplary embodiment of the present invention, any ofthe antibodies provided herein may comprise a CDRH3 sequence which maybe 100% identical to SEQ ID NO.: 178.

These heavy chain variable domains may comprise a CDRH1 sequence atleast 80% identical to SEQ ID NO.:182, a CDRH2 sequence at least 80%identical to SEQ ID NO.:183 and a CDRH3 sequence at least 80% identicalto SEQ ID NO.:184.

In an exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRH1 sequence which may be atleast 90% identical to SEQ ID NO.: 182.

In another exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRH1 sequence which may be100% identical to SEQ ID NO.: 182.

In yet another exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRH2 sequence which may be atleast 90% identical to SEQ ID NO.: 183.

In a further exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRH2 sequence which may be100% identical to SEQ ID NO.: 183.

In yet a further exemplary embodiment of the present invention, any ofthe antibodies provided herein may comprise a CDRH3 sequence which maybe at least 90% identical to SEQ ID NO.: 184.

In an additional exemplary embodiment of the present invention, any ofthe antibodies provided herein may comprise a CDRH3 sequence which maybe 100% identical to SEQ ID NO.: 184.

These heavy chain variable domains may comprise a CDRH1 sequence atleast 80% identical to SEQ ID NO.:188, a CDRH2 sequence at least 80%identical to SEQ ID NO.:189 and a CDRH3 sequence at least 80% identicalto SEQ ID NO.:190.

In an exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRH1 sequence which may be atleast 90% identical to SEQ ID NO.: 188.

In another exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRH1 sequence which may be100% identical to SEQ ID NO.: 188.

In yet another exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRH2 sequence which may be atleast 90% identical to SEQ ID NO.: 189.

In a further exemplary embodiment of the present invention, any of theantibodies provided herein may comprise a CDRH2 sequence which may be100% identical to SEQ ID NO.: 189.

In yet a further exemplary embodiment of the present invention, any ofthe antibodies provided herein may comprise a CDRH3 sequence which maybe at least 90% identical to SEQ ID NO.: 190.

In an additional exemplary embodiment of the present invention, any ofthe antibodies provided herein may comprise a CDRH3 sequence which maybe 100% identical to SEQ ID NO.: 190.

Production of the Antibodies in Cells

The antibodies that are disclosed herein can be made by a variety ofmethods familiar to those skilled in the art, such as hybridomamethodology or by recombinant DNA methods.

In an exemplary embodiment of the invention, the antibodies may beproduced by the conventional hybridoma technology, where a mouse isimmunized with an antigen, spleen cells isolated and fused with myelomacells lacking HGPRT expression and hybrid cells selected byhypoxanthine, aminopterin and thymine (HAT) containing media.

In an additional exemplary embodiment of the invention, the antibodiesmay be produced by recombinant DNA methods.

In order to express the antibodies, nucleotide sequences able to encodeany one of a light and heavy immunoglobulin chains described herein maybe inserted into an expression vector, i.e., a vector that contains theelements for transcriptional and translational control of the insertedcoding sequence in a particular host. These elements may includeregulatory sequences, such as enhancers, constitutive and induciblepromoters, and 5′ and 3′ un-translated regions. Methods that are wellknown to those skilled in the art may be used to construct suchexpression vectors. These methods include in vitro recombinant DNAtechniques, synthetic techniques, and in vivo genetic recombination.

A variety of expression vector/host cell systems known to those of skillin the art may be utilized to express a polypeptide or RNA derived fromnucleotide sequences able to encode any one of a light and heavyimmunoglobulin chains described herein. These include, but are notlimited to, microorganisms such as bacteria transformed with recombinantbacteriophage, plasmid, or cosmid DNA expression vectors; yeasttransformed with yeast expression vectors; insect cell systems infectedwith baculovirus vectors; plant cell systems transformed with viral orbacterial expression vectors; or animal cell systems. For long-termproduction of recombinant proteins in mammalian systems, stableexpression in cell lines may be effected. For example, nucleotidesequences able to encode any one of a light and heavy immunoglobulinchains described herein may be transformed into cell lines usingexpression vectors that may contain viral origins of replication and/orendogenous expression elements and a selectable or visible marker geneon the same or on a separate vector. The invention is not to be limitedby the vector or host cell employed. In certain embodiments of thepresent invention, the nucleotide sequences able to encode any one of alight and heavy immunoglobulin chains described herein may each beligated into a separate expression vector and each chain expressedseparately. In another embodiment, both the light and heavy chains ableto encode any one of a light and heavy immunoglobulin chains describedherein may be ligated into a single expression vector and expressedsimultaneously.

Alternatively, RNA and/or polypeptide may be expressed from a vectorcomprising nucleotide sequences able to encode any one of a light andheavy immunoglobulin chains described herein using an in vitrotranscription system or a coupled in vitro transcription/translationsystem respectively.

In general, host cells that contain nucleotide sequences able to encodeany one of a light and heavy immunoglobulin chains described hereinand/or that express a polypeptide encoded by the nucleotide sequencesable to encode any one of a light and heavy immunoglobulin chainsdescribed herein, or a portion thereof, may be identified by a varietyof procedures known to those of skill in the art. These proceduresinclude, but are not limited to, DNA/DNA or DNA/RNA hybridizations, PCRamplification, and protein bioassay or immunoassay techniques thatinclude membrane, solution, or chip based technologies for the detectionand/or quantification of nucleic acid or amino acid sequences.Immunological methods for detecting and measuring the expression ofpolypeptides using either specific polyclonal or monoclonal antibodiesare known in the art. Examples of such techniques include enzyme-linkedimmunosorbent assays (ELISAs), radioimmunoassays (RIAs), andfluorescence activated cell sorting (FACS). Those of skill in the artmay readily adapt these methodologies to the present invention.

Host cells comprising nucleotide sequences able to encode any one of alight and heavy immunoglobulin chains described herein may thus becultured under conditions for the transcription of the corresponding RNA(mRNA, siRNA, shRNA etc.) and/or the expression of the polypeptide fromcell culture. The polypeptide produced by a cell may be secreted or maybe retained intracellularly depending on the sequence and/or the vectorused. In an exemplary embodiment, expression vectors containingnucleotide sequences able to encode any one of a light and heavyimmunoglobulin chains described herein may be designed to contain signalsequences that direct secretion of the polypeptide through a prokaryoticor eukaryotic cell membrane.

Due to the inherent degeneracy of the genetic code, other DNA sequencesthat encode the same, substantially the same or a functionallyequivalent amino acid sequence may be produced and used, for example, toexpress a polypeptide encoded by nucleotide sequences able to encode anyone of a light and heavy immunoglobulin chains described herein. Thenucleotide sequences of the present invention may be engineered usingmethods generally known in the art in order to alter the nucleotidesequences for a variety of purposes including, but not limited to,modification of the cloning, processing, and/or expression of the geneproduct. DNA shuffling by random fragmentation and PCR reassembly ofgene fragments and synthetic oligonucleotides may be used to engineerthe nucleotide sequences. For example, oligonucleotide-mediatedsite-directed mutagenesis may be used to introduce mutations that createnew restriction sites, alter glycosylation patterns, change codonpreference, produce splice variants, and so forth.

In addition, a host cell strain may be chosen for its ability tomodulate expression of the inserted sequences or to process theexpressed polypeptide in the desired fashion. Such modifications of thepolypeptide include, but are not limited to, acetylation, carboxylation,glycosylation, phosphorylation, lipidation, and acylation. In anexemplary embodiment, antibodies that contain particular glycosylationstructures or patterns may be desired. Post-translational processing,which cleaves a “prepro” form of the polypeptide, may also be used tospecify protein targeting, folding, and/or activity. Different hostcells that have specific cellular machinery and characteristicmechanisms for post-translational activities (e.g., CHO, HeLa, MDCK,HEK293, and W138) are available commercially and from the American TypeCulture Collection (ATCC) and may be chosen to ensure the correctmodification and processing of the expressed polypeptide.

Since hybridoma cells are hybrid mouse cells, they are strictly used toproduce murine antibodies. It is clear that the glycosyl side chains ofsuch murine antibodies might significantly differ from the glycosylationpattern observed in human cells. Differences in phosphorylation patternbetween human cells and hybridomas might also have an impact on theactivity of the antibody. Furthermore, administration of murineantibodies to human usually induces an anti-antibody immune responsethat could potentially neutralize any of the biological activity thatthe murine antibody might have.

In order to minimize recognition of murine antibodies by the humanimmune system or for improving the biological activity of the antibodiesin human, murine antibodies are advantageously converted into partially(e.g., chimeric) or fully humanized antibodies. Recombinant form of thelight chain and heavy chain of the (partially or fully) humanizedantibody may thus be introduced into a mammalian expression system otherthan hybridoma cells (such as 293 cells, CHO or else). Mammalianexpression system may procure the advantage of having a resultingglycosylation pattern that is closer to that of naturally occurringhuman form of the antibodies.

For example, in the case of lytic IgG1 antibodies, the properglycosylation of the immunoglobulin chains is necessary for effectorfunctions. These biological functions of IgG1 monoclonal antibodiesinclude antibody-dependent cell cytotoxicity (ADCC) andcomplement-dependent cytotoxicity (CDC), both of which will be greatlyinfluenced by the type of glycosyl side chains that are grafted to theamino acids during expression in mammalian cells.

In addition, optimized mammalian cell expression systems will oftensecrete significantly a greater amounts of antibodies compared tohybridomas. Therefore, there is a practical and probably economicalreason for adopting human cells for production.

Those of skill in the art will readily appreciate that natural,modified, or recombinant nucleic acid sequences may be ligated to aheterologous sequence resulting in translation of a fusion polypeptidecontaining heterologous polypeptide moieties in any of theaforementioned host systems. Such heterologous polypeptide moieties mayfacilitate purification of fusion polypeptides using commerciallyavailable affinity matrices. Such moieties include, but are not limitedto, glutathione S-transferase (GST), maltose binding protein,thioredoxin, calmodulin binding peptide, 6-His (His), FLAG, c-myc,hemaglutinin (HA), and antibody epitopes such as monoclonal antibodyepitopes.

In yet a further aspect, the present invention relates to apolynucleotide which may comprise a nucleotide sequence encoding afusion protein. The fusion protein may comprise a fusion partner (e.g.,HA, Fc, etc.) fused to the polypeptide (e.g., complete light chain,complete heavy chain, variable regions, CDRs etc.) described herein.

Those of skill in the art will also readily recognize that the nucleicacid and polypeptide sequences may be synthesized, in whole or in part,using chemical or enzymatic methods well known in the art. For example,peptide synthesis may be performed using various solid-phase techniquesand machines such as the ABI 431A Peptide synthesizer (PE Biosystems)may be used to automate synthesis. If desired, the amino acid sequencemay be altered during synthesis and/or combined with sequences fromother proteins to produce a variant protein.

Antibody Conjugates

Although it is not always necessary, for detection or therapeuticpurposes, the antibody or antigen binding fragment of the presentinvention may be conjugated with a detectable moiety (i.e., fordetection or diagnostic purposes) or with a therapeutic moiety (fortherapeutic purposes).

For detection purposes, an unconjugated antibody (primary antibody) maybe used for binding to the antigen and a secondary antibody carrying adetectable moiety and capable of binding to the primary antibody may beadded. However, as indicated above, the anti-SIGLEC 15 antibody may beconjugated with a detectable label and as such a secondary antibody maynot be necessary,

A “detectable moiety” is a moiety detectable by spectroscopic,photochemical, biochemical, immunochemical, chemical and/or otherphysical means. A detectable moiety may be coupled either directlyand/or indirectly (for example via a linkage, such as, withoutlimitation, a DOTA or NHS linkage) to antibodies and antigen bindingfragments thereof of the present invention using methods well known inthe art. A wide variety of detectable moieties may be used, with thechoice depending on the sensitivity required, ease of conjugation,stability requirements and available instrumentation. A suitabledetectable moiety include, but is not limited to, a fluorescent label, aradioactive label (for example, without limitation, ¹²⁵I, In¹¹¹, Tc⁹⁹,I¹³¹ and including positron emitting isotopes for PET scanner etc), anuclear magnetic resonance active label, a luminescent label, achemiluminescent label, a chromophore label, an enzyme label (forexample and without limitation horseradish peroxidase, alkalinephosphatase, etc.), quantum dots and/or a nanoparticle. Detectablemoiety may cause and/or produce a detectable signal thereby allowing fora signal from the detectable moiety to be detected.

In another exemplary embodiment of the invention, the antibody orantigen binding fragment thereof may be coupled (modified) with atherapeutic moiety (e.g., drug, cytotoxic moiety).

In some instances, for therapeutic purposes, an unconjugated antibodymay by itself be capable of sequestering the antigen, may block animportant interaction between the antigen and another binding partner,may recruit effector cells, etc. However, as indicated above, theantibody may be conjugated with a therapeutic moiety.

In an exemplary embodiment, the antibodies and antigen binding fragmentsmay comprise a chemotherapeutic or cytotoxic agent. For example, theantibody and antigen binding fragments may be conjugated to thechemotherapeutic or cytotoxic agent. Such chemotherapeutic or cytotoxicagents include, but are not limited to, Yttrium-90, Scandium-47,Rhenium-186, Iodine-131, Iodine-125, and many others recognized by thoseskilled in the art (e.g., lutetium (e.g., Lu¹⁷⁷), bismuth (e.g., Bi²¹³),copper (e.g., Cu⁶⁷)). In other instances, the chemotherapeutic orcytotoxic agent may be comprised of, among others known to those skilledin the art, 5-fluorouracil, adriamycin, irinotecan, taxanes, pseudomonasendotoxin, ricin and other toxins.

Alternatively, in order to carry out the methods of the presentinvention and as known in the art, the antibody or antigen bindingfragment of the present invention (conjugated or not) may be used incombination with a second molecule (e.g., a secondary antibody, etc.)which is able to specifically bind to the antibody or antigen bindingfragment of the present invention and which may carry a desirabledetectable, diagnostic or therapeutic moiety.

Pharmaceutical Compositions of the Antibodies and their Use

Pharmaceutical compositions of the antibodies (conjugated or not) arealso encompassed by the present invention. The pharmaceuticalcomposition may comprise an antibody or an antigen binding fragment andmay also contain a pharmaceutically acceptable carrier.

Other aspects of the invention relate to a composition which maycomprise the antibody or antigen binding fragment described herein and acarrier.

Yet other aspects of the invention relate to the use of the isolatedantibody or antigen binding fragment described herein in the treatmentor diagnosis of bone diseases or cancer.

In addition to the active ingredients, a pharmaceutical composition maycontain pharmaceutically acceptable carriers comprising water, PBS, saltsolutions, gelatins, oils, alcohols, and other excipients andauxiliaries that facilitate processing of the active compounds intopreparations that may be used pharmaceutically. In other instances, suchpreparations may be sterilized.

As used herein, “pharmaceutical composition” usually comprisestherapeutically effective amounts of the agent together withpharmaceutically acceptable diluents, preservatives, solubilizers,emulsifiers, adjuvant and/or carriers. A “therapeutically effectiveamount” as used herein refers to that amount which provides atherapeutic effect for a given condition and administration regimen.Such compositions are liquids or lyophilized or otherwise driedformulations and include diluents of various buffer content (e.g.,Tris-HCl., acetate, phosphate), pH and ionic strength, additives such asalbumin or gelatin to prevent absorption to surfaces, detergents (e.g.,Tween 20, Tween 80, Pluronic F68, bile acid salts). Solubilizing agents(e.g., glycerol, polyethylene glycerol), anti-oxidants (e.g., ascorbicacid, sodium metabisulfite), preservatives (e.g., thimerosal, benzylalcohol, parabens), bulking substances or tonicity modifiers (e.g.,lactose, mannitol), covalent attachment of polymers such as polyethyleneglycol to the protein, complexation with metal ions, or incorporation ofthe material into or onto particulate preparations of polymericcompounds such as polylactic acid, polyglycolic acid, hydrogels, etc, oronto liposomes, microemulsions, micelles, unilamellar or multilamellarvesicles, erythrocyte ghosts, or spheroplasts. Such compositions willinfluence the physical state, solubility, stability, rate of in vivorelease, and rate of in vivo clearance. Controlled or sustained releasecompositions include formulation in lipophilic depots (e.g., fattyacids, waxes, oils). Also comprehended by the invention are particulatecompositions coated with polymers (e.g., poloxamers or poloxamines).Other embodiments of the compositions of the invention incorporateparticulate forms protective coatings, protease inhibitors or permeationenhancers for various routes of administration, including parenteral,pulmonary, nasal, oral, vaginal, rectal routes. In one embodiment thepharmaceutical composition is administered parenterally, paracancerally,transmucosally, transdermally, intramuscularly, intravenously,intradermally, subcutaneously, intraperitonealy, intraventricularly,intracranially and intratumorally.

Further, as used herein “pharmaceutically acceptable carrier” or“pharmaceutical carrier” are known in the art and include, but are notlimited to, 0.01-0.1 M or 0.05 M phosphate buffer or 0.8% saline.Additionally, such pharmaceutically acceptable carriers may be aqueousor non-aqueous solutions, suspensions, and emulsions. Examples ofnon-aqueous solvents are propylene glycol, polyethylene glycol,vegetable oils such as olive oil, and injectable organic esters such asethyl oleate. Aqueous carriers include water, alcoholic/aqueoussolutions, emulsions or suspensions, including saline and bufferedmedia. Parenteral vehicles include sodium chloride solution, Ringer'sdextrose, dextrose and sodium chloride, lactated Ringer's or fixed oils.Intravenous vehicles include fluid and nutrient replenishers,electrolyte replenishers such as those based on Ringer's dextrose, andthe like. Preservatives and other additives may also be present, suchas, for example, antimicrobials, antioxidants, collating agents, inertgases and the like.

For any compound, the therapeutically effective dose may be estimatedinitially either in cell culture assays or in animal models such asmice, rats, rabbits, dogs, or pigs. An animal model may also be used todetermine the concentration range and route of administration. Suchinformation may then be used to determine useful doses and routes foradministration in humans. These techniques are well known to one skilledin the art and a therapeutically effective dose refers to that amount ofactive ingredient that ameliorates the symptoms or condition.Therapeutic efficacy and toxicity may be determined by standardpharmaceutical procedures in cell cultures or with experimental animals,such as by calculating and contrasting the ED₅₀ (the dosetherapeutically effective in 50% of the population) and LD₅₀ (the doselethal to 50% of the population) statistics. Any of the therapeuticcompositions described above may be applied to any subject in need ofsuch therapy, including, but not limited to, mammals such as dogs, cats,cows, horses, rabbits, monkeys, and humans.

The pharmaceutical compositions utilized in this invention may beadministered by any number of routes including, but not limited to,oral, intravenous, intramuscular, intra-arterial, intramedullary,intrathecal, intraventricular, transdermal, subcutaneous,intraperitoneal, intranasal, enteral, topical, sublingual, or rectalmeans.

The term “treatment” for purposes of this disclosure refers to boththerapeutic treatment and prophylactic or preventative measures, whereinthe object is to prevent or slow down (lessen) the targeted pathologiccondition or disorder. Those in need of treatment include those alreadywith the disorder as well as those prone to have the disorder or thosein whom the disorder is to be prevented.

The antibodies or antigen binding fragments may have therapeutic uses inthe treatment of various bone loss or cancer. In an exemplaryembodiment, the antibodies or fragments may have therapeutic uses inbone loss associated with bone diseases such as conditions where thereis an increase in the bone degradative activity of osteoclasts. Incertain instances, the antibodies or antigen binding fragments mayinteract with cells that express SEQ ID NO:2 and induce an immunologicalreaction by mediating ADCC. In other instances, the antibodies andfragments may block the interaction of SEQ ID NO:2 with its proteinpartners.

The anti-Siglec-15 antibodies or antigen binding fragments may havetherapeutic uses in the treatment of bone loss in the context of variousbone-related diseases, including but not limited to osteoporosis,osteopenia, osteomalacia, hyperparathyroidism, hypothyroidism,hyperthyroidism, hypogonadism, thyrotoxicosis, systemic mastocytosis,adult hypophosphatasia, hyperadrenocorticism, osteogenesis imperfecta,Paget's disease, Cushing's disease/syndrome, Turner syndrome, Gaucherdisease, Ehlers-Danlos syndrome, Marfan's syndrome, Menkes' syndrome,Fanconi's syndrome, multiple myeloma, hypercalcemia, hypocalcemia,arthritides, periodontal disease, rickets (including vitamin Ddependent, type I and II, and x-linked hypophosphatemic rickets),fibrogenesis imperfecta ossium, osteosclerotic disorders such aspycnodysostosis and damage caused by macrophage-mediated inflammatoryprocesses. In the preferred embodiment, the antibodies and fragmentshave therapeutic uses in conditions where severe bone loss prevails, inparticular metastatic cancer to the bone. In certain instances, theanti-Siglec-15 antibodies and fragments may interact with cells, such asosteoclasts, that express Siglec-15. In other instances, theanti-Siglec-15 antibodies and fragments may block the interaction ofSiglec-15 with its protein partners.

The anti-Siglec-15 antibodies and antigen binding fragments thereof mayhave therapeutic uses in the treatment of cancer or bone loss caused byor associated with various bone remodelling disorders. In particular,the anti-Siglec-15 antibodies and immunologically functional fragmentstherein have therapeutic uses in conditions where osteoclasts arehyperactive and contribute to the degradation of the bone surface. Incertain instances, the anti-Siglec-15 antibodies and antigen bindingfragment thereof may be administered concurrently in combination withother treatments given for the same condition. As such, the antibodiesmay be administered with anti-resorptives (e.g., bisphosphonates) thatare known to those skilled in the art. Additionally, the antibodies maybe administered with anti-mitotics (e.g., taxanes), platinum-basedagents (e.g., cisplatin), DNA damaging agents (e.g. Doxorubicin), andother cytotoxic therapies that are known to those skilled in the art. Inother instances, the anti-Siglec-15 antibodies and immunologicallyfunctional fragments therein may be administered with other therapeuticantibodies. These include, but are not limited to, antibodies thattarget RANKL, EGFR, CD-20, and Her2.

In certain instances, the antibodies and antigen binding fragmentstherein may be administered concurrently in combination with othertreatments given for the same condition. As such, the antibodies may beadministered with anti-mitotics (e.g., taxanes), platinum-based agents(e.g., cisplatin), DNA damaging agents (e.g. Doxorubicin), and otheranti-cancer therapies that are known to those skilled in the art. Inother instances, the antibodies and antigen binding fragments thereinmay be administered with other therapeutic antibodies. These include,but are not limited to, antibodies that target EGFR, CD-20, and Her2.

The present invention relates in a further aspect thereof to a methodfor inhibiting the growth of a SEQ ID NO:2-expressing cell or of SEQ IDNO:2 variant-expressing cell, the method which may comprise contactingthe cell with an effective amount of the antibody or antigen bindingfragment described herein.

The present invention also encompasses method of treating cancer or boneloss or inhibiting the growth of a SEQ ID NO:2 expressing cells or ofSEQ ID NO:2 variant-expressing cell in a mammal, the method may compriseadministering the antibody or antigen binding fragment described hereinto a mammal in need.

The present invention also provides a method for inhibiting the growthof a cancer cell selected from the group consisting of ovarian cancercells, renal cancer cells, cancer cells of the central nervous system,prostate cancer cells, melanoma cells, breast cancer cells, lung cancercells or colon cancer cells. The method may comprise providing thecancer cell with a nucleic acid capable of impairing the expression of apolypeptide at least 80% identical to SEQ ID NO.:2 or having a region atleast 80% identical to amino acids 20 to 259 or to amino acids 49 to 165of SEQ ID NO.:2. The cancer cell may express a polypeptide at least 80%identical to SEQ ID NO.:2 or having a region at least 80% identical toamino acids 20 to 259 or to amino acids 49 to 165 of SEQ ID NO.:2.

In accordance with the present invention, the nucleic acid may be, forexample, a siRNA or an antisense.

The present invention also encompasses method of detecting cancer orbone loss or detecting a SEQ ID NO:2-expressing cell or a SEQ ID NO:2variant-expressing cell in a mammal, the method may compriseadministering the antibody or antigen binding fragment described hereinto a mammal in need.

The present invention relates in another aspect thereof to a method fordetecting a SEQ ID NO:2-expressing cell or a SEQ ID NO:2variant-expressing cell, the method may comprise contacting the cellwith an antibody or antigen binding fragment described herein anddetecting a complex formed by the antibody and the SEQ IDNO:2-expressing cell or the SEQ ID NO:2 variant-expressing cell.

Another aspect of the invention relates to a method for detecting SEQ IDNO:2, a variant having at least 80% sequence identity with amino acids20-259 or with amino acids 49-165 of SEQ ID NO:2, the method maycomprise contacting a cell expressing SEQ ID NO:2 or the variant or asample (biopsy, serum, plasma, urine etc.) comprising or suspected ofcomprising SEQ ID NO:2 or the variant with the antibody or antigenbinding fragments described herein and measuring binding.

The binding of an antibody to an antigen will cause an increase in theexpected molecular weight of the antigen. A physical change thereforeoccurs upon specific binding of the antibody or antigen binding fragmentand the antigen.

Such changes may be detected using, for example, electrophoresisfollowed by Western blot and coloration of the gel or blot, massspectrometry, HPLC coupled with a computer or else. Apparatus capable ofcomputing a shift in molecular weight are known in the art and includefor example, Phosphorimager™.

When the antibody comprises for example a detectable label, theantigen-antibody complex may be detected by the fluorescence emitted bythe label, radiation emission of the label, enzymatic activity of alabel provided with its substrate or else.

Detection and/or measurement of binding between an antibody or antigenbinding fragment and an antigen may be performed by various methodsknown in the art. Binding between an antibody or antigen bindingfragment and an antigen may be monitored with an apparatus capable ofdetecting the signal emitted by the detectable label (radiationemission, fluorescence, color change etc.). Such apparatus provides datawhich indicates that binding as occurred and may also provide indicationas to the amount of antibody bound to the antigen. The apparatus(usually coupled with a computer) may also be capable of calculating thedifference between a background signal (e.g., signal obtained in theabsence of antigen-antibody binding) or background noise and the signalobtained upon specific antibody-antigen binding. Such apparatuses maythus provide the user with indications and conclusions as to whether theantigen has been detected or not.

The sample may originate from a mammal (e.g., a human) which may havecancer or bone disease or may be suspected of having cancer or a bonedisease or may experience bone loss or may be subject of experiencingbone loss. The sample may be a tissue sample obtained from the mammal ora cell culture supernatant.

In accordance with the invention the sample may be a serum sample, aplasma sample, a blood sample or ascitic fluid obtained from the mammal.The antibody or antigen binding fragment described herein mayadvantageously detect SEQ ID NO:2.

The method may comprise quantifying the complex formed by the antibodyor antigen binding fragment bound to SEQ ID NO:2 or to the SEQ ID NO:2variant.

The antibody or antigen binding fragment of the present invention maymore particularly be used in the detection, diagnosis or treatment ofbone disease or cancer.

Additional aspects of the invention relates to kits which may includeone or more container containing one or more antibodies or antigenbinding fragments described herein.

Nucleic Acids, Vectors and Cells

Antibodies are usually made in cells allowing expression of the lightchain and heavy chain expressed from a vector(s) comprising a nucleicacid sequence encoding the light chain and heavy chain.

The present therefore encompasses nucleic acids capable of encoding anyof the CDRs (including CDR variants), light chain variable domains(including light chain variable domain variants), heavy chain variabledomains (including heavy chain variable domain variants), light chains(including light chain variants), heavy chains (including heavy chainvariants) described herein.

Exemplary embodiments of nucleic acids of the present invention includenucleic acids encoding a light chain variable domain comprising:

-   -   a. a CDRL1 sequence selected from the group consisting of SEQ ID        NO:69, SEQ ID NO:75, SEQ ID NO:81, SEQ ID NO:87, SEQ ID NO:93,        SEQ ID NO:99, SEQ ID NO:105 and SEQ ID NO:111;    -   b. a CDRL2 sequence selected from the group consisting of SEQ ID        NO:70, SEQ ID NO:76. SEQ ID NO:82. SEQ ID NO:88. SEQ ID NO:94,        SEQ ID NO:100, SEQ ID NO:106 and SEQ ID NO:112, and/or;    -   c. a CDRL3 sequence selected from the group consisting of SEQ ID        NO:71, SEQ ID NO:77, SEQ ID NO:83, SEQ ID NO:89, SEQ ID NO:95,        SEQ ID NO:101, SEQ ID NO:107 and SEQ ID NO:113.

In accordance with the present invention, the nucleic acid may encode alight chain variable domain which may comprise at least two CDRs of aCDRL1, a CDRL2 or a CDRL3.

Also in accordance with the present invention, the nucleic acid mayencode a light chain variable domain which may comprise one CDRL1, oneCDRL2 and one CDRL3.

The present invention also relates to a nucleic acid encoding a heavychain variable domain comprising:

-   -   a. a CDRH1 sequence selected from the group consisting of SEQ ID        NO:72, SEQ ID NO:78, SEQ ID NO:84, SEQ ID NO:90, SEQ ID NO:96,        SEQ ID NO:102, SEQ ID NO:108 and SEQ ID NO:114;    -   b. a CDRH2 sequence selected from the group consisting of SEQ ID        NO:73, SEQ ID NO:79, SEQ ID NO:85, SEQ ID NO:91, SEQ ID NO:97,        SEQ ID NO:103, SEQ ID NO:109 and SEQ ID NO:115, and/or;    -   c. a CDRH3 sequence selected from the group consisting of SEQ ID        NO:74, SEQ ID NO:80, SEQ ID NO:86, SEQ ID NO:92, SEQ ID NO:98,        SEQ ID NO:104, SEQ ID NO:110 and SEQ ID NO:116.

In a further aspect, the present invention provides a nucleic acidencoding a light chain variable domain which may comprise:

-   -   a) a CDRL1 sequence selected from the group consisting of SEQ ID        NO: 153, SEQ ID NO.:154, SEQ ID NO.:84, SEQ ID NO.:96 and SEQ ID        NO.:102;    -   b) a CDRL2 sequence selected from the group consisting of SEQ ID        NO: 149, SEQ ID NO.:150, SEQ ID NO.:76, SEQ ID NO.:82 and SEQ ID        NO.:106, or;    -   c) a CDRL3 sequence selected from the group consisting of SEQ ID        NO: 151, SEQ ID NO.:152, SEQ ID NO.:77, SEQ ID NO.:83, SEQ ID        NO.:95, SEQ ID NO.:107 and SEQ ID NO.:152.

In yet a further aspect, the present invention provides a nucleic acidencoding a heavy chain variable domain which may comprise:

-   -   a) a CDRH1 sequence selected from the group consisting of SEQ ID        NO: 153, SEQ ID NO.:154, SEQ ID NO.:84, SEQ ID NO.:96 and SEQ ID        NO.:102;    -   b) a CDRH2 sequence selected from the group consisting of SEQ ID        NO: 155, SEQ ID NO.:156, SEQ ID NO.:157, SEQ ID NO.:73, SEQ ID        NO.:79, SEQ ID NO.:85, SEQ ID NO.:97, SEQ ID NO.:103 and SEQ ID        NO.:109, or;    -   c) a CDRH3 sequence selected from the group consisting of SEQ ID        NO: 158, SEQ ID NO.:74, SEQ ID NO.:98, SEQ ID NO.:104, SEQ ID        NO.:110 and SEQ ID NO.:116.

In accordance with the present invention, the nucleic acid may encode aheavy chain variable domain which may comprise at least two CDRs of aCDRH1, a CDRH2 or a CDRH3.

In accordance with the present invention, the nucleic acid may encode aheavy chain variable domain which may comprise one CDRH1, one CDRH2 andone CDRH3.

Also encompassed by the present invention are nucleic acids encodingantibody variants having at least one conservative amino acidsubstitution.

In accordance with the present invention, the nucleic acid may encode aCDR comprising at least one conservative amino acid substitution.

In accordance with the present invention, the nucleic acid may encode aCDR comprising at least one conservative amino acid substitution in atleast two of the CDRs.

In accordance with the present invention, the nucleic acid may encode aCDR comprising at least one conservative amino acid substitution in the3 CDRs.

In accordance with the present invention, the nucleic acid may encode aCDR comprising at least two conservative amino acid substitution in atleast one of the CDRs.

In accordance with the present invention, the nucleic acid may encode aCDR comprising at least two conservative amino acid substitution in atleast two of the CDRs.

In accordance with the present invention, the nucleic acid may encode aCDR comprising at least two conservative amino acid substitution in the3 CDRs.

Other aspects of the invention relate to a nucleic acid encoding a lightchain variable domain having at least 70% (including at least 80%)sequence identity to a sequence selected from the group consisting ofSEQ ID NO:37, SEQ ID NO.:41, SEQ ID NO.:45, SEQ ID NO.:49, SEQ IDNO.:53, SEQ ID NO.:57, SEQ ID NO.:61 and SEQ ID NO.:65.

Yet other aspects of the invention relate to a nucleic acid encoding aheavy chain variable domain having at least 70% (including at least 80%)sequence identity to a sequence selected from the group consisting ofSEQ ID NO.:39, SEQ ID NO.:43, SEQ ID NO.:47, SEQ ID NO.:51, SEQ IDNO.:55, SEQ ID NO.:59, SEQ ID NO.:63 and SEQ ID NO.:67.

In yet another aspect, the present invention relates to a vectorcomprising the nucleic acid described herein.

In accordance with the present invention, the vector may be anexpression vector.

Vector that contains the elements for transcriptional and translationalcontrol of the inserted coding sequence in a particular host are knownin the art. These elements may include regulatory sequences, such asenhancers, constitutive and inducible promoters, and 5′ and 3′un-translated regions. Methods that are well known to those skilled inthe art may be used to construct such expression vectors. These methodsinclude in vitro recombinant DNA techniques, synthetic techniques, andin vivo genetic recombination.

In another aspect the present invention relates to an isolated cellwhich may comprise the nucleic acid described herein.

The isolated cell may comprise a nucleic acid encoding a light chainvariable domain and a nucleic acid encoding a heavy chain variabledomain either on separate vectors or on the same vector. The isolatedcell may also comprise a nucleic acid encoding a light chain and anucleic acid encoding a heavy chain either on separate vectors or on thesame vector.

In accordance with the present invention, the cell may be capable ofexpressing, assembling and/or secreting an antibody or antigen bindingfragment thereof.

In another aspect, the present invention provides a cell which maycomprise and/or may express the antibody described herein.

In accordance with the invention, the cell may comprise a nucleic acidencoding a light chain variable domain and a nucleic acid encoding aheavy chain variable domain.

The cell may be capable of expressing, assembling and/or secreting anantibody or antigen binding fragment thereof.

The examples below are presented to further outline details of thepresent invention.

Exemplary Embodiments of Screening Assay

In an additional aspect the present invention provides methods ofidentifying a compound capable of inhibiting the growth of ovariancancer cells, renal cancer cells, cancer cells of the central nervoussystem, prostate cancer cells, melanoma cells, breast cancer cells, lungcancer cells or colon cancer cells. The method may comprise providing apolypeptide comprising a region at least 80% identical to amino acids 20to 259 of SEQ ID NO.:2 or a cell expressing said polypeptide with acandidate compound and measuring the activity or expression of thepolypeptide. A reduced activity or expression of the polypeptide maypositively identify a suitable inhibitory compound.

In accordance with the present invention, the candidate compound mayspecifically bind to the polypeptide.

In accordance with the present invention, the candidate compound may be,for example, an antibody or an antigen binding fragment.

In accordance with the present invention, the candidate compound may be,for example, a siRNA or an antisense.

Other types of assay may be carried out without departing from the scopeof the invention.

EXAMPLES Example 1

This example describes the pattern of expression of the Siglec-15 genein osteoclasts and human tissue RNA samples

One of the most promising genes identified was termed AB-0326, whichencodes the cell surface type I membrane protein, Siglec-15. Thiscandidate was first isolated from a human osteoclast library and asimilar RANKL-dependent upregulation was also confirmed in primary mouseosteoclasts as well as the mouse RAW 264.7 cells compared to precursorcells by RT-PCR (Sooknanan et al. 2007). The tissue expression profileof Siglec-15 was assessed to determine the specificity of expression, acriteria that was imposed on all targets that were chosen forvalidation. Peripheral blood mononuclear cells (PBMNCs) were obtainedfrom 6 human donors and cultured in osteoclast differentiation medium(MCS-F and RANKL) for at least 14 days. Total RNA was isolated fromprecursors cells (no RANKL treatment (FIG. 1, top panel, −) or atintermediate time intervals (FIG. 1, top panel,

). One microgram of each RNA sample was converted to single-strandedcDNA using Thermoscript reverse transcriptase (Invitrogen, Burlington,ON) according to the manufacturer's instructions, diluted 200-fold, andused in a PCR reaction previously optimized to specifically amplify afragment of the Siglec-15 transcript. The sequences of theoligonucleotides used in the PCR reaction are shown in SEQ ID NOS: 117and 118. As shown in FIG. 1 top panel, differentiating osteoclasts), theSiglec-15 transcript was either expressed at much lower level in theprecursors cells compared to the differentiating osteoclasts. Inaddition, the level of Siglec-15 transcript increased as thedifferentiation progressed. By comparison, a known osteoclast markergene, cathepsin K (CATK in FIG. 1, differentiating osteoclasts) was alsoupregulated during osteoclast differentiation. The oligonucleotides usedto amplify the CATK message are displayed in SEQ ID NOS: 119 and 120. Asa control, PCR reactions were conducted on the same samples with primersthat specifically amplify the housekeeping geneglyceraldehyde-3-phosphate dehydrogenase (GAPDH, see lower panel of FIG.1, differentiating osteoclasts). The sequences of the GAPDH-specificprimers used in the PCR reaction are shown in SEQ ID NOS: 121 and 122.This latter reaction demonstrates that an equal amount of starting RNAwas present in each sample. Total RNA from human normal tissues waspurchased from a commercial vendor (Clontech, Mountain View, Calif.) Asshown in FIG. 1 (upper panel, Normal tissues), Siglec-15 was weaklydetected in a single tissue (lung, lane 9) and completely absent fromall other tissue samples. This underscores the strength of theApplicant's discovery approach in its ability to identify targets thatare highly restricted to differentiating osteoclasts. The lane numbersin FIG. 1 correspond to the following tissues: The lanes correspond tothe following tissues: lane 1, adrenal; 2, breast; 3, jejunum; 4,trachea; 5, liver; 6, placenta; 7, aorta; 8, brain; 9, lung; 10, adrenalcortex; 11, esophagus; 12, colon; 13, ovary; 14, kidney; 15, prostate;16, thymus; 17, skeletal muscle; 18, vena cava; 19, stomach; 20, smallintestine; 21, heart; 22, fallopian tube; 23, spleen; 24, bladder; 25,cervix; 26, pancreas; 27, ileum; 28, duodenum; 29, thyroid; 30,testicle; the blank lanes between lanes 10 and 11 and lanes 20 and 21represent negative controls (no cDNA). Our results indicate thatSiglec-15 is upregulated in differentiating osteoclasts, absent fromvirtually all normal human tissues and suggest that an antibody againstSiglec-15 would interact significantly less with non-target tissues.

An additional expression profiling study was performed to determine theexpression of Siglec-15 in cancer indications. One skilled in the artwill recognize that the antibodies described in this invention mighthave utilities in cancer if it was found that the Siglec-15 gene wasexpressed in these types of indications. To address this, the PCR-basedmethod was adapted to determine the expression pattern of the Siglec-15transcript in cancer cell lines isolated from nine types of cancer. Thecancer types represented by the cell lines are leukemia, central nervoussystem, breast, colon, lung, melanoma, ovarian, prostate, and renalcancer (see Table 4). These RNA samples were obtained from theDevelopmental Therapeutics Program at the NCl/NIH. Using the same RAMPRNA samples that was amplified from the total RNA samples obtained fromthe NCl, 500 ng of RNA was converted to single-stranded cDNA asdescribed above. The cDNA reaction was diluted so that 1/200 of thereaction was used for each PCR experiment. PCR was conducted in 96-wellplates using Hot-Start Taq Polymerase from Qiagen (Mississauga, ON) in aDNA Engine Tetrad from MJ Research. Half of the reaction mixture wasloaded on a 1.2% agarose/ethidium bromide gel and the ampliconsvisualized with UV light. To verify that equal quantities of RNA wasused in each reaction, the level of RNA was monitored with GAPDHexpression.

TABLE 4 List of cancer cell lines from the NCI-60 panel Cell line Cancertype K-562 leukemia MOLT-4 leukemia CCRF-CEM leukemia RPMI-8226 leukemiaHL-60(TB) leukemia SR leukemia SF-268 CNS SF-295 CNS SF-539 CNS SNB-19CNS SNB-75 CNS U251 CNS BT-549 breast HS 578T breast MCF7 breastNCI/ADR-RES breast MDA-MB-231 breast MDA-MB-435 breast T-47D breast COLO205 colon HCC-2998 colon HCT-116 colon HCT-15 colon HT29 colon KM12colon SW-620 colon A549/ATCC non-small cell lung EKVX non-small celllung HOP-62 non-small cell lung HOP-92 non-small cell lung NCI-H322Mnon-small cell lung NCI-H226 non-small cell lung NCI-H23 non-small celllung NCI-H460 non-small cell lung NCI-H522 non-small cell lung LOX IMVImelanoma M14 melanoma MALME-3M melanoma SK-MEL-2 melanoma SK-MEL-28melanoma SK-MEL-5 melanoma UACC-257 melanoma UACC-62 melanoma IGROV-1ovarian OVCAR-3 ovarian OVCAR-4 ovarian OVCAR-5 ovarian OVCAR-8 ovarianSK-OV-3 ovarian DU-145 prostate PC-3 prostate 786-O renal A498 renalACHN renal CAKI-1 renal

As shown in FIG. 2, Siglec-15 was found to be expressed in severalcancer types, in particular ovarian cancer, renal cancer, cancer of thecentral nervous system, and prostate cancer. In fact, Siglec-15 wasdetected in almost every cancer indication represented by these sampleswith the exception of leukemia. This result suggests that antibodiesagainst Siglec-15 might have uses in cancer diseases.

The antibodies described in Example 2 (see below) may also be used fordetection of Siglec-15 in cell lysates by immunoblotting. The entireopen reading frame of human Siglec-15 cDNA was cloned into a mammalianexpression vector downstream of a CMV promoter (pcDNA-Siglec-15). Thisconstruct, or a control empty vector which does not encode Siglec-15,were transfected into A375 melanoma cells, which express low endogenouslevels of Siglec-15 protein. A pool of stable transfectants was isolatedby selection with G418. Cell lysates from Siglec-15-transfected (+) andcontrol (−) A375 cells were analysed by immunoblotting with monoclonalantibody E6. As shown in FIG. 13A, the antibody detects a single band of35 kDa in the Siglec-15-transfected cells, but not in control cells.This closely matches the predicted molecular weight of Siglec-15 (35.62kDa), based on the primary amino acid sequence(http://www.bioinformatics.org/sms/prot_mw.html). Lysates were alsoanalyzed by immunoblotting with an anti-β-actin antibody to demonstratethat similar total amounts of lysates were loaded in each lane. Thisresult demonstrates that, by immunoblotting, antibody E6 recognizes, ina highly specific manner, overexpressed Siglec-15 in lysates from cellstransfected with cloned Siglec-15 cDNA.

To confirm that the increased Siglec-15 mRNA levels in differentiatedhuman PBMNC (FIG. 1) correspond to an increase in Siglec-15 proteinlevels, lysates were prepared from human PBMNC treated with MCSF alone(non-differentiated, C) or MCSF and RANKL (differentiated, Δ) (FIG.13B). Lysates were also prepared from RAW 264.7 cells left untreated(non-differentiated, C) or treated with RANKL (differentiated, Δ) (FIG.13C). RAW 264.7 cells were shown previously to upregulate Siglec-15 mRNAlevels upon induction of osteoclast differentiation by RANKL (Sooknanan,2007). Analysis of these lysates by immunoblotting with antibody E9demonstrates that, as predicted by RT-PCR studies, there is a dramaticincrease in Siglec-15 protein levels both in PBMNC and RAW 264.7 cellsupon differentiation into osteoclasts (FIGS. 13B and 13C).

RT-PCR analysis of mRNA from the NCl60 panel (FIG. 2) indicated that aparticularly high proportion of CNS-derived cancer cell lines expressSiglec-15, while a recent microarray study found a small set of cancercell lines, including the U87 glioblastoma line that is not part of theNCl60 panel, that express very high levels of Siglec-15 mRNA(Shankavaram, 2007). Therefore, we tested whether endogenous expressionof Siglec-15 protein could be detected in U87 cells. Indeed, a proteinthe size of Siglec-15 is detected by immunoblotting of U87 cell lysates.To confirm the identity of this protein, U87 cells were transfected witha pool of small interfering RNAs (siRNAs) targeting Siglec-15 (SIGLEC15siGENOME SMARTpool, Dharmacon) (+) or with a control, non-targetingsiRNA pool (−, FIG. 13D) and allowed to grow for 72 h before cell lysis.Consistent with its identification as Siglec-15, treatment with thetargeted siRNA resulted in reduced expression of this protein comparedto the non-targeted control (FIG. 13D). To examine whether Siglec-15 isfound at the cell surface in cancer cells, we analyzed the siRNA-treatedU87 cells by flow cytometry. Living cells were placed on ice and stainedwith Siglec-15 antibody E9 (see Example 2) or an isotype controlantibody, under conditions which allow antibody binding to extracellularbut not intracellular antigens. Treatment with the targeted siRNAresulted in reduced binding of antibody E9 but had no effect on bindingof the control antibody (FIG. 13E). Together, these results demonstratethat Siglec-15 may be expressed in cancer cells, and that it isaccessible for antibody binding at the cell surface.

Example 2

This example provides details pertaining to the family of monoclonalantibodies that bind to Siglec-15.

To generate monoclonal antibodies, recombinant human Siglec-15 wasproduced in 293E cells using the large-scale transient transfectiontechnology (Durocher et al., 2002; Durocher, 2004). A cDNA encodingamino acids 20-259 of SEQ ID NO:2 (see SEQ ID NO:123) was amplified byPCR using a forward primer that incorporated a BamHI restriction site(SEQ ID NO:124) and a reverse primer that incorporated a NotIrestriction site (SEQ ID NO:125). The resulting PCR product was digestedwith BamHI and NotI and the fragment was ligated into the expressionvector pYD5 (SEQ ID NO:126) that was similarly digested with the samerestriction enzymes to create a vector called pYD5-0326. The pYD5expression plasmid contains the coding sequence for the human Fc domainthat allows fusion proteins to be generated as well as the sequenceencoding the IgG1 signal peptide to allow the secretion of the fusionprotein into the culture medium. For each milliliter of cells, onemicrogram of the expression vector, called pYD5-0326₂₀₋₂₅₉, wastransfected in 2936E cells grown in suspension to a density of 1.5-2.0million cells/ml. The transfection reagent used was polyethylenimine(PEI), (linear, MW 25,000, Cat#23966 Polysciences, Inc., Warrington,Pa.) which was included at a DNA:PEI ratio of 1:3. Growth of the cellswas continued for 5 days after which the culture medium was harvestedfor purification of the recombinant Fc-0326₂₀₋₂₅₉ fusion protein. Theprotein was purified using Protein-A agarose as instructed by themanufacturer (Sigma-Aldrich Canada Ltd., Oakville, ON). A representativepolyacrylamide gel showing a sample of the purified Fc-0326₂₀₋₂₅₉(indicated as Fc-Siglec-15₂₀₋₂₅₉) is shown in FIG. 3.

The antibodies that bind Siglec-15 were generated using the Biositephage display technology. A detailed description of the technology andthe methods for generating these antibodies can be found in the U.S.Pat. No. 6,057,098. Briefly, the technology utilizes stringent panningof phage libraries that display the antigen binding fragments (Fabs).After a several rounds of panning, a library, termed the Omniclonal, wasobtained that was enriched for recombinant Fabs containing light andheavy chain variable regions that bound to Siglec-15 with very highaffinity and specificity. From this library, more precisely designatedOmniclonal AL0025Z1, 96 individual recombinant monoclonal Fabs wereprepared from E. coli and tested for Siglec-15 binding.

To measure the relative binding of each individual monoclonal antibody,recombinant human Fc-Siglec-15₂₀₋₂₅₉ was produced in 293E cells usingthe large-scale transient transfection technology (Durocher et al.,2002; Durocher, 2004). The 96-well master plate of monoclonalpreparations contained different concentrations of purifiedanti-Siglec-15 Fabs in each well. A second stock master plate wasprepared by diluting the Fabs to a final concentration of 10 μg/ml fromwhich all subsequent dilutions were performed for ELISA measurements. Tocarry out the binding of Fc-Siglec-15 to the monoclonal preparations,the Fc-Siglec-15₂₀₋₂₅₉ was biotinylated with NHS-biotin (Pierce,Rockford, Ill.) and 10 ng/well was coated in a streptavidin 96-wellplate. One nanogram of each Fab monoclonal preparation was added to eachwell and incubated at room temperature for 30 minutes. Bound antibodywas detected with HRP-conjugated mouse anti-kappa light chain antibodyin the presence of TMB liquid substrate (Sigma-Aldrich Canada Ltd.,Oakville, ON) and readings were conducted at 450 nm in microtiter platereader. As shown in FIG. 4A, a total of 53 (highlighted dark grey)monoclonal antibodies displayed significant binding in this assay (>0.2arbitrary OD₄₅₀ units). The antibodies were purposely diluted to 1ng/well to accentuate the binding of those antibodies with the mostaffinity for Siglec-15. Since the antibodies were generated using a Fcfusion protein, the monoclonals were also tested in an ELISA usingbiotinylated Fc domain only. As shown in FIG. 4B, 17 antibodiesinteracted with the Fc moiety of the Fc-Siglec-15₂₀₋₂₅₉ (highlightedlight grey). The values presented in bold (see FIG. 4) represent theexemplary antibodies 25A1, 25B4, 25B8, 25C1, 25D8, 25E5, 25E6, and 25E9.These data also revealed that the binding of the antibodies varied fromwell to well indicating that they exhibited different affinities forSiglec-15.

The applicant noted that the antibody or antigen binding fragment of thepresent invention may bind efficiently to the antigen, in fact it wasfound that 1 ng of antibody is capable of binding to less than 500 ng ofSEQ ID NO.:2.

The specificity of these antibodies for Siglec-15 was assessed bytesting their binding to two other members of the Siglec family, CD33and Siglec-2. CD33 (GeneBank™ accession No. NM_(—)001772.3) is theprototype of the CD33-related family of Siglecs: among human proteins,these Siglecs share the highest amino acid sequence similarity withSiglec-15 (around 29% sequence identity between their two respectiveN-terminal Ig-like domains). Siglec-2 (GeneBank™ accession No.NM_(—)001771.3) is less similar (23% sequence identity), but likeSiglec-15 and unlike most other Siglecs, it has a marked preference forbinding α2-6-linked sialic acid conjugates (Angata 2007, Blixt 2003).Sequences comprising the V-set and N-terminal C2-set Ig-like domains ofSiglec-2 and CD33 (corresponding to the region of Siglec-15 used as theantigen for antibody production) were cloned from a human PBMNC cDNAlibrary into the pYD5 vector. Supernatants from 293-6E cells transfectedwith these constructs, as wells as from non-transfected 293-6E cells orthose transfected with pYD5-Siglec-15 or pYD5 empty vector, wereanalyzed by immunoblotting with an anti-Fc antibody to evaluateexpression levels (FIG. 14A). Transfection of these constructs resultedin expression of Fc-tagged proteins of the expected size (FIG. 14A).Aliquots of these supernatants were adsorbed onto PVDF by vacuum dotblotting (Bio-dot apparatus, Bio-Rad), and binding of representativeSiglec-15 monoclonal antibodies was evaluated (Western blots were notused because many antibodies react only with the native, non-denaturedform of Siglec-15). As controls, anti-Fc and anti-Siglec-15 omniclonalantibodies reacted with all four Fc-tagged proteins (FIG. 14B). Incontrast, monoclonal antibodies D8 and E9 show no detectable binding toFc alone, Siglec-2 or CD33, indicating that they are highly specific forSiglec-15.

Example 3

This example discloses the methods used to convert the Fabs into fullIgG2 chimeric monoclonal antibodies. A scheme of the methodology ispresented in FIG. 5.

In order to conduct in vitro and in vivo studies to validate thebiological function of the antigen the light and heavy chain variableregions contained in the Fabs was transferred to full antibodyscaffolds, to generate mouse-human chimeric IgG2s. The expressionvectors for both the light and heavy immunoglobulin chains wereconstructed such that i) the original bacterial signal peptide sequencesupstream of the Fab expression vectors were replaced by mammalian signalpeptides and ii) the light and heavy chain constant regions in the mouseantibodies were replaced with human constant regions. The methods toaccomplish this transfer utilized standard molecular biology techniquesthat are familiar to those skilled in the art. A brief overview of themethodology is described here (see FIG. 5).

Light Chain Expression Vector—

an existing mammalian expression plasmid, called pTTVH8G (Durocher etal., 2002), designed to be used in a 293E transient transfection systemwas modified to accommodate the mouse light chain variable region. Theresulting mouse-human chimeric light chain contained a mouse variableregion followed by the human kappa constant domain. The cDNA sequenceencoding the human kappa constant domain was amplified by PCR withprimers OGS1773 and OGS1774 (SEQ ID NOS:127 and 128, respectively). Thenucleotide sequence and the corresponding amino acid sequence for thehuman kappa constant region are shown in SEQ ID NOS:129 and 130,respectively. The resulting 321 base pair PCR product was ligated intopTTVH8G immediately downstream of the signal peptide sequence of humanVEGF A (NM_(—)003376). This cloning step also positioned uniquerestriction endonuclease sites that permitted the precise positioning ofthe cDNAs encoding the mouse light chain variable regions. The sequenceof the final expression plasmid, called pTTVK1, is shown in SEQ IDNO:131. Based on the sequences disclosed in Table 2, PCR primersspecific for the light chain variable regions of antibodies 25A1, 25B4,25B8, 25C1, 25D8, 25E5, 25E6, and 25E9 (SEQ ID NOS:37, 41, 45, 49, 53,57, 61, and 65, respectively) were designed that incorporated, at their5′-end, a sequence identical to the last 20 base pairs of the VEGF Asignal peptide. The sequences of these primers are shown in SEQ IDNO:132 for 25A1; SEQ ID NO:133 for 25B4, 25B8, 25C1, 25D8, and 25E9; SEQID NO:134 for 25E5, and SEQ ID NO:135 for 25E6, respectively. The samereverse primer was used to amplify all four light chain variable regionssince the extreme 3′-ends were identical. This primer (SEQ ID NO:136)incorporated, at its 3′-end, a sequence identical to the first 20 basepairs of the human kappa constant domain. Both the PCR fragments and thedigested pTTVK1 were treated with the 3′-5′ exonuclease activity of T4DNA polymerase resulting in complimentary ends that were joined byannealing. The annealing reactions were transformed into competent E.coli and the expression plasmids were verified by sequencing to ensurethat the mouse light chain variable regions were properly inserted intothe pTTVK1 expression vector. Those skilled in the art will readilyrecognize that the method used for construction of the light chainexpression plasmids applies to all anti-Siglec-15 antibodies containedin the original Fab library.

Heavy Chain Expression Vector—

the expression vector that produced the heavy chain immunoglobulins wasdesigned in a similar manner to the pTTVK1 described above forproduction of the light chain immunoglobulins. In the case of thechimeric anti-Siglec-15 antibodies, IgG2 isotype was required which isthe preferred type for stable, blocking antibodies. To this end, theconstant regions (CH1, CH2, and CH3) of the human IgG2 immunoglobulinwere amplified and ligated into a pre-existing IgG1 expression vectorand the detailed methods are described herein. Plasmid pYD11 (Durocheret al., 2002), which contains the human IgGK signal peptide sequence aswell as the CH2 and CH3 regions of the human Fc domain of IgG1, wasmodified by ligating the cDNA sequence encoding the human constant CH1region. PCR primers OGS1769 and OGS1770 (SEQ ID NOS:137 and 138),designed to contain unique restriction endonuclease sites, were used toamplify the human IgG1 CH1 region containing the nucleotide sequence andcorresponding amino acid sequence shown in SEQ ID NOS:139 and 140.Following ligation of the 309 base pair fragment of human CH1immediately downstream of the IgGK signal peptide sequence, theresulting plasmid was digested with the restriction enzymes ApaI andNsiI. These enzymes that digest both the constant IgG1 and IgG2 cDNAs inexactly the same positions that permits the IgG1 constant sequence to bereplaced by the human IgG2 sequence in the expression vector. The cDNAencoding the human IgG2 constant domains was obtained from acommercially available source (Open Biosystems, Huntsville, Ala.). Thefinal plasmid used to express the IgG2 immunoglobulin heavy chain wasdesignated pYD19 and the sequence is shown in SEQ ID NO:141. When aselected heavy chain variable region is ligated into this vector, theresulting plasmid encodes a full IgG2 heavy chain immunoglobulin withhuman constant regions. Based on the sequences disclosed in Table 2, PCRprimers specific for the heavy chain variable regions of antibodies25A1, 25B4, 25B8, 25C1, 25D8, 25E5, 25E6, and 25E9 (SEQ ID NOS:39, 43,47, 51, 55, 59, 63, and 67, respectively) were designed thatincorporated, at their 5′-end, a sequence identical to the last 20 basepairs of the IgGK signal peptide. The sequences of these primers areshown in SEQ ID NOS:142 for 25A1; SEQ ID NO:143 for 24B4 and 25D8; SEQID NO:144 for 25B8, 25C1, and 25E9; SEQ ID NO:145 for 25E5; and SEQ IDNO:146 for 25E6, respectively. The same reverse primer was used toamplify all four heavy chain variable regions since the extreme 3′-endswere identical. This primer (SEQ ID NO:147) incorporated, at its 3′-end,a sequence identical to the first 20 base pairs of the human CH1constant domain. Both the PCR fragments and the digested pYD19 weretreated with the 3′-5′ exonuclease activity of T4 DNA polymeraseresulting in complimentary ends that were joined by annealing. Theannealing reactions were transformed into competent E. coli and theexpression plasmids were verified by sequencing to ensure that the mouseheavy chain variable regions were properly inserted into the pYD19expression vector. Those skilled in the art will readily recognize thatthe method used for construction of the heavy chain expression plasmidsapplies to all anti-Siglec-15 antibodies contained in the original Fablibrary.

Expression of Human IgG2s in 293E Cells—

The expression vectors prepared above that encoded the light and heavychain immunoglobulins were expressed in 293E cells using the transienttransfection system (Durocher et al., 2002). By virtue of the signalpeptides incorporated at the amino-termini of both immunoglobulinchains, the mature IgG2 was harvested from the serum-free culture mediumof the cells. The methods used for co-transfecting the light and heavychain expression vectors were described herein. For each milliliter ofcells, one microgram of a combination of both the light and heavy chainexpression plasmids was transfected in 293E cells grown in suspension toa density of 1.5-2.0 million cells/ml. The ratio of light to heavy chainplasmid was optimized in order to achieve the most yield of antibody inthe tissue culture medium and it was found to be 9:1 (L:H). Thetransfection reagent used was polyethylenimine (PEI), (linear, MW25,000, Cat#23966 Polysciences, Inc., Warrington, Pa.) which wasincluded at a DNA:PEI ratio of 1:3. Growth of the cells was continuedfor 5 days after which the culture medium was harvested for purificationof the IgG2 chimeric monoclonal antibodies. The protein was purifiedusing Protein-A agarose as instructed by the manufacturer (Sigma-AldrichCanada Ltd., Oakville, ON).

To determine the relative binding affinity of selected monoclonals moreaccurately, increasing concentration of the Fabs was incubated withbiotinylated Fc-Siglec-15₂₀₋₂₅₉. Ten nanograms of biotinylatedFc-Siglec-15₂₀₋₂₅₉ was coated in streptavidin microtiter plates andincreasing amounts of either Fabs or the chimeric IgG2 monoclonals 25B4,25B8, 25C1, 25D8, 25E6, and 25E9 were added as indicated in FIG. 6. Asdepicted in FIG. 6, the binding of the 25B4, 25B8, 25C1, 25D8, 25E6, and25E9 chimeric IgG2 monoclonal antibodies was very similar to the Fabs.This result shows that the transposition of the variable domains fromthe mouse Fabs into a human IgG2 backbone did not significantly affectthe capacity of the light and heavy chain variable regions to conferSiglec-15 binding.

Example 4

This example describes the use of anti-Siglec-15 antibodies forinhibiting the differentiation of osteoclasts.

Human PBMNCs (AllCells, Emoryville, Calif.) were placed in theappropriate culture medium for 24 h at 37 C in a 5% CO₂ atmosphere. Thecells were seeded in 96-well plates at a cell density of 100,000cells/ml and treated with increasing concentration (0.01 μg/ml-100μg/ml) of anti-Siglec-15 IgG2 chimeric monoclonal antibodies in thepresence of 35 ng/ml M-CSF and 30 ng/ml RANKL. Undifferentiatedprecursor cells were treated only with M-CSF, The control wells weretreated with a non-Siglec-15 binding IgG2. The cells were fixed, stainedfor TRAP, and multinucleated cells counted and photographed(magnification 40×). As depicted in FIG. 7, mAbs targeting Siglec-15could efficiently inhibit the differentiation of human osteoclasts in adose-dependent manner. Inhibition of osteoclast differentiation wasobserved to varying extents with every exemplary Siglec-15 antibody thatwas tested but the most active monoclonals were 25B8, 25E6, and 25E9.Cells treated with a control chimeric IgG2 were not inhibited (see lowerright panels in FIG. 8, Control IgG2). This result is in completeagreement with the experiments disclosed by Sooknanan (Sooknanan et al.,2007) that showed that knockdown of Siglec-15 expression by RNAinterference caused inhibition of human osteoclast differentiation.

The biological function of differentiated osteoclasts is to resorb boneand thus the activity of osteoclasts should also be inhibited byantibodies that target Siglec-15. To test this, human PBMNCs were seededon synthetic calcium phosphate substrate discs (BD BioCoat™ Osteologic™MultiTest Slides) and cultured in similar conditions as described above.The precursor cells were treated with M-CSF and RANKL in the presence ofeither a control isotype IgG or the 25D8 or 25E9 anti-Siglec-15antibodies. The antibodies were at a concentration of 1 μg/ml or 10μg/ml. Once fully matured osteoclasts were present in the controluntreated wells. The cells were scarped off the discs and the remainingbone substrate was stained using a standard von Kossa stain whichrenders the calcium mineral brown. As shown in FIG. 12, the wellscontaining undifferentiated osteoclasts (upper left panel, M-CSF) showedno evidence of degradation of the substrate which appears as white spotson the surface (degradation pits). As expected, the cells treated withRANKL had evidence of significant degradation and the surface containedmany pits (lower left panel, M-CSF+RANKL). Similarly, the osteoclaststreated with the control IgG could also degrade the bone substrate whichdemonstrated that these control antibodies did not inhibit osteoclastactivity non-specifically. When the differentiating osteoclasts weretreated with the anti-Siglec-15 antibodies, the 25E9 candidateefficiently inhibited bone degradation in this assay (FIG. 12, rightpanels). By contrast, the 25D8 antibody did not inhibit degradation inthis assays (see middle-right panels of FIG. 12). Taken together, theseresults (FIG. 7 and FIG. 12) demonstrate that antibodies againstSiglec-15 inhibit osteoclast differentiation and bone degradationactivity.

In a parallel experiment, mouse PBMNCs were treated in a similar manner.As depicted in FIG. 8, anti-Siglec-15 chimeric antibodies could inhibitthe differentiation of mouse osteoclasts as exemplified by the chimericmAbs designated 25B8, 25E6, and 25D8. This result confirms that themonoclonal antibodies that were generated against the human orthologueof Siglec-15 are cross-reactive against the mouse Siglec-15 protein aswell. This was experimentally verified using an ELISA. A fragment of themouse Siglec-15 cDNA was amplified corresponding to amino acids 21-256using oligonucleotides containing the sequences shown in SEQ ID NOS: 159and 160. This PCR fragment was ligated into the pYD5 expression vectoras was described for the human Siglec-15 fragment for expression in293-6E cells. The recombinant Fc-mouse Siglec-15 was purified usingProtein-A affinity chromatography.

An exemplary anti-Siglec-15 monoclonal Fab designated 25C8 was incubatedwith either Fc-human(h)Siglec-15-20-259 or Fc-mouse(m)Siglec-15₂₁₋₂₅₆.The results (see FIG. 9) indicate that the binding activity of theantibodies that were generated against the human Siglec-15 alsocross-react with the mouse orthologue of Siglec-15.

The results described above clearly demonstrate the importance ofSiglec-15 in osteoclastogenesis. Attenuation of Siglec-15 expression inosteoclast precursor cells results in cells that are highly impaired intheir ability to form multinucleated mature osteoclasts. Thus, targetingSiglec-15 with an inhibitor, in particular a therapeutic monoclonalantibody, would prove to be a very selective way to target those cellsthat are directly responsible for bone degradation during acutemetastatic bone cancer or chronic osteoporosis.

Example 5

This example pertains to the ability of anti-Siglec-15 antibodies toblock binding of Siglec-15 to sialic acid (SA) conjugates.

The formation of sialylated glycoproteins is required for properosteoclastogenesis (Takahata et al., 2007). Siglec-15 binds sialic acid,and this binding is dependent on the amino acid residue R143 (Angata2007). One mechanism by which Siglec-15 antibodies inhibit osteoclastformation could involve interference with their target's sialicacid-binding function due to interactions with an epitope encompassingR143. To examine this possibility, we performed an ELISA-based assay totest the ability of Siglec-15 antibodies to block binding of recombinantFc-Siglec-15 to Neu5Aca2-6-GalNAc-PAA-Biotin (Glycotech, Rockville,Md.), which is a preferred, sialic acid-containing binding partner ofSiglec-15 (Angata 2007). Fc-Siglec-15 was immobilized on a ProteinA-coated microtiter plate, and different Siglec-15 antibodies were thenapplied. After incubation and removal of unbound antibody,Neu5Aca2-6-GalNAc-PAA-Biotin was added. This biotinylated probe shouldform a complex with Siglec-15 only if an antibody is not blocking thesialic acid binding site. The presence of the biotinylated probe wasdetected using streptavidin-HRP by standard methods. As shown in FIG.15, anti-Siglec-15 omniclonal and 25D8 antibodies inhibit sialic acidbinding compared to a non-targeting, control antibody. Antibody E6 alsohas a clear, but less pronounced effect. Antibody E9 has little effect,indicating that its epitope does not overlap with the sialic acidbinding site. Addition of a control antibody (FIG. 15, see ctl IgG2) didnot prevent the binding of sialic acid moiety to Siglec-15. The methodwas highly dependent on the presence of Siglec-15 since no binding wasdetected when only the Fc was coated in the plates nor was there anybinding when the SA was omitted (FIG. 15, see no SA, Fc+SA, and Fconly). Together, these results demonstrate that the Siglec-15 monoclonalantibodies can interfere, to varying extents, with the sialic acidbinding function of Siglec-15 likely due to interactions near R143. Thisproperty could be important for their effects on osteoclastogenesis.

CITED REFERENCES

-   Frost H. M., 1964 Dymanics of Bone Remodelling. In: Bone    Biodynamics, Little and Brown, Boston, Mass., USA pp. 315;-   Baron, R., Anatomy and Biology of Bone Matrix and Cellular Elements,    In: Primer on the Metabolic Bone Diseases and Disorders of Mineral    Metabolism, Fifth Edition 2003, American Society for Bone and    Mineral Research, Washington D.C., pp. 1-8;-   Jilka, R. L. et al., “Increased Osteoclast Development After    Estrogen Loss: Mediation by Interleukin-6”, Science 257: 88-91    (1992).-   Poli, V. et al., “Interleukin-6 deficient mice are protected from    bone loss caused by estrogen depletion”, EMBO J 13: 1189-1196    (1994).-   Srivastava, S. et al., “Estrogen Blocks M-CSF Gene Expression and    Osteoclast Formation by Regulating Phosphorylation of Egr-1 and Its    Interaction with Sp-1”, J Clin Invest 102: 1850-1859 (1998).-   de Vernejoul, M. C., “Dynamics of Bone Remodelling: Biochemical and    Pathophysiological Basis”, Eur J Clin Chem Clin Biochem 34: 729-734    (1996).-   McMillan, S. J. and P. R. Crocker, “CD33-related sialic-acid-binding    immunoglobulin-like lectins in health and disease”, Carbohydr Res,    343(12): p. 2050-6 (2008).-   Crocker, P. R., J. C. Paulson, and A. Varki, Siglecs and their roles    in the immune system. Nat Rev Immunol, 7(4): p. 255-66 (2007).-   Angata, T., et al., Siglec-15: an immune system Siglec conserved    throughout vertebrate evolution. Glycobiology, 17(8): p. 838-46    (2007).-   Sooknanan, R. R., “Polynucleotides and polypeptide sequences    involved in the process of bone remodelling”, PCT/CA2007/000210    (2007).-   Takahata, M., et al., Sialylation of cell surface glycoconjugates is    essential for osteoclastogenesis. Bone, 41(1): p. 77-86 (2007).-   Ellis, G. K. et al., “Randomized Trial of Denosumab in Patients    Receiving Adjuvant Aromatase Inhibitors for Nonmetastatic Breast    Cancer”, J Clin Oncol 26: 4875-4882 (2008).-   Buechler J, Valkirs G, Gray J. “Polyvalent display libraries.” U.S.    Pat. No. 6,057,098 (2000).-   Durocher Y, Kamen A, Perret S, Pham P L. “Enhanced production of    recombinant proteins by transient transfection of suspension-growing    mammalian cells.” Canadian patent application No. CA 2446185 (2002).-   Durocher Y. “Expression vectors for enhanced transient gene    expression and mammalian cells expressing them.” U.S. patent    application No. 60/662,392 (2004).-   Shankavaram, U. T. et al., “Transcript and protein expression    profiles of the NCI-60 cancer panel: an integromic microarray    study”, Mol Cancer Ther 6: 820-832 (2007).-   Blixt O. et al., “Sialoside specificity of the siglec family    assessed using novel multivalent probes”, J Biol Chem, 278,    31007-31019.

SEQUENCE LISTINGS SEQ ID NO: 1ATGGAAAAGTCCATCTGGCTGCTGGCCTGCTTGGCGTGGGTTCTCCCGACAGGCTCATTTGTGAGAACTAAAATAGATACTACGGAGAACTTGCTCAACACAGAGGTGCACAGCTCGCCAGCGCAGCGCTGGTCCATGCAGGTGCCACCCGAGGTGAGCGCGGAGGCAGGCGACGCGGCAGTGCTGCCCTGCACCTTCACGCACCCGCACCGCCACTACGACGGGCCGCTGACGGCCATCTGGCGCGCGGGCGAGCCCTATGCGGGCCCGCAGGTGTTCCGCTGCGCTGCGGCGCGGGGCAGCGAGCTCTGCCAGACGGCGCTGAGCCTGCACGGCCGCTTCCGGCTGCTGGGCAACCCGCGCCGCAACGACCTCTCGCTGCGCGTCGAGCGCCTCGCCCTGGCTGACGACCGCCGCTACTTCTGCCGCGTCGAGTTCGCCGGCGACGTCCATGACCGCTACGAGAGCCGCCACGGCGTCCGGCTGCACGTGACAGCCGCGCCGCGGATCGTCAACATCTCGGTGCTGCCCAGTCCGGCTCACGCCTTCCGCGCGCTCTGCACTGCCGAAGGGGAGCCGCCGCCCGCCCTCGCCTGGTCCGGCCCGGCCCTGGGCAACAGCTTGGCAGCCGTGCGGAGCCCGCGTGAGGGTCACGGCCACCTAGTGACCGCCGAACTGCCCGCACTGACCCATGACGGCCGCTACACGTGTACGGCCGCCAACAGCCTGGGCCGCTCCGAGGCCAGCGTCTACCTGTTCCGCTTCCATGGCGCCAGCGGGGCCTCGACGGTCGCCCTCCTGCTCGGCGCTCTCGGCTTCAAGGCGCTGCTGCTGCTCGGGGTCCTGGCCGCCCGCGCTGCCCGCCGCCGCCCAGAGCATCTGGACACCCCGGACACCCCACCACGGTCCCAGGCCCAGGAGTCCAATTATGAAAATTTGAGCCAGATGAACCCCCGGAGCCCACCAGCCACCATGTGCTCACCGTGA SEQ ID NO: 2MEKSIWLLACLAWVLPTGSFVRTKIDTTENLLNTEVHSSPAQRWSMQVPPEVSAEAGDAAVLPCTFTHPHRHYDGPLTAIWRAGEPYAGPQVFRCAAARGSELCQTALSLHGRFRLLGNPRRNDLSLRVERLALADDRRYFCRVEFAGDVHDRYESRHGVRLHVTAAPRIVNISVLPSPAHAFRALCTAEGEPPPALAWSGPALGNSLAAVRSPREGHGHLVTAELPALTHDGRYTCTAANSLGRSEASVYLFRFHGASGASTVALLLGALGFKALLLLGVLAARAARRRPEHLDTPDTPPRSQAQESNYENLSQMNPRSPPATMCSP SEQ ID NO: 3ATGGAGGGGTCCCTCCAACTCCTGGCCTGCTTGGCCTGTGTGCTCCAGATGGGATCCCTTGTGAAAACTAGAAGAGACGCTTCGGGGGATCTGCTCAACACAGAGGCGCACAGTGCCCCGGCGCAGCGCTGGTCCATGCAGGTGCCCGCGGAGGTGAACGCGGAGGCTGGCGACGCGGCGGTGCTGCCCTGCACCTTCACGCACCCGCACCGCCACTACGACGGGCCGCTGACGGCCATCTGGCGCTCGGGCGAGCCGTACGCGGGCCCGCAGGTGTTCCGCTGCACCGCGGCGCCGGGCAGCGAGCTGTGCCAGACGGCGCTGAGCCTGCACGGCCGCTTCCGCCTGCTGGGCAACCCGCGCCGCAACGACCTGTCCCTGCGCGTCGAGCGCCTCGCCCTGGCGGACAGCGGCCGCTACTTCTGCCGCGTGGAGTTCACCGGCGACGCCCACGATCGCTATGAGAGTCGCCATGGGGTCCGTCTGCGCGTGACTGCAGCTGCGCCGCGGATCGTCAACATCTCGGTGCTGCCGGGCCCCGCGCACGCCTTCCGCGCGCTCTGCACCGCCGAGGGGGAGCCCCCGCCCGCCCTCGCCTGGTCGGGTCCCGCCCCAGGCAACAGCTCCGCTGCCCTGCAGGGCCAGGGTCACGGCTACCAGGTGACCGCCGAGTTGCCCGCGCTGACCCGCGACGGCCGCTACACGTGCACGGCGGCCAATAGCCTGGGCCGCGCCGAGGCCAGCGTCTACCTGTTCCGCTTCCACGGCGCCCCCGGAACCTCGACCCTAGCGCTCCTGCTGGGCGCGCTGGGCCTCAAGGCCTTGCTGCTGCTTGGCATTCTGGGAGCGCGTGCCACCCGACGCCGACTAGATCACCTGGTCCCCCAGGACACCCCTCCACGGTCTCAGGCTCAGGAGTCCAATTATGAAAATTTGAGCCAGATGAGTCCTCCAGGCCACCAGCTGCCACGTGTTTGCTGTGAGGAACTCCTCAGCCATCACCATCTAGTCATTCACCATGAGAAATAA SEQ ID NO: 4MEGSLQLLACLACVLQMGSLVKTRRDASGDLLNTEAHSAPAQRWSMQVPAEVNAEAGDAAVLPCTFTHPHRHYDGPLTAIWRSGEPYAGPQVFRCTAAPGSELCQTALSLHGRFRLLGNPRRNDLSLRVERLALADSGRYFCRVEFTGDAHDRYESRHGVRLRVTAAAPRIVNISVLPGPAHAFRALCTAEGEPPPALAWSGPAPGNSSAALQGQGHGYQVTAELPALTRDGRYTCTAANSLGRAEASVYLFRFHGAPGTSTLALLLGALGLKALLLLGILGARATRRRLDHLVPQDTPPRSQAQESNYENLSQMSPPGHQLPRVCCEELLSHHHLVIHHEK SEQ ID NO: 5GAAAATGTGCTCACCCAGTCTCCAGCAATCATGTCTGCATCTCCAGGGGAGAAGGTCACCATATCCTGCAGTGCCAGCTCAAGTGTAAGTTACATGTACTGGTACCAGCAGAAGCCAGGATCCTCCCCCAAACCCTGGATTTATCGCACATCCAACCTGGCTTCTGGAGTCCCTGCTCGCTTCAGTGGCAGTGGGTCTGGGACCTCTTACTCTCTCACAATCAGCAGCATGGAGGCTGAAGATGCTGCCACTTATTACTGCCAGCAGTGGAGTAGTAACCCACTCACGTTCGGTGCTGGGACCAAGCTGGAGCTGAAACGGGCTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAG SEQ ID NO: 6ENVLTQSPAIMSASPGEKVTISCSASSSVSYMYWYQQKPGSSPKPWIYRTSNLASGVPARFSGSGSGTSYSLTISSMEAEDAATYYCQQWSSNPLTFGAGTKLELKVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQ ID NO: 7GAGGTCCAGCTGCAACAATCTGGGACTGAGCTGGTGAGGCCTGGGTCCTCAGTGAAGATTTCCTGCAAGGCTTCTGGCTACACCTTCACCAGGTACTGGATGGACTGGGTGAAGCAGAGGCCTGGACAAGGCCTTGAGTGGATCGGAGAGATTGATCCTTCTGATAGTTATACTAACTACAATCAAAAGTTCAAGGGCAAGGCCACATTGACTGTAGATAAATTCTCCAGAACAGCCTATATGGAACTCAGCAGCCTGACATCTGAGGACTCTGCGGTCTATTACTGTGCAAGATCGGGGGCCTACTCTAGTGACTATAGTTACGACGGGTTTGCTTACTGGGGCCAAGGGACTCTGGTCACTGTCTCTGCAGCCTCAACGAAGGGCCCATCGGTCTTCCCCCTGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCTCTGACCAGCGGCGTGCACACCTTCCCAGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAACTTCGGCACCCAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGACAGTTGAGCGCAAATGTTGTGTCGAGTGCCCACCGTGCCCAGCACCACCTGTGGCAGGACCGTCAGTCTTCCGCTTCCCCCCAAAACCCAAGGACACCCGCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGATGTGAGCCACGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCACGGGAGGAGCAGTTCAACAGCACGTTCCGTGTGGTCAGCGTCCTCACCGTTGTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGGCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAACCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACACCTCCCATGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAATGA SEQ ID NO: 8EVQLQQSGTELVRPGSSVKISCKASGYTFTRYWMDWVKQRPGQGLEWIGEIDPSDSYTNYNQKFKGKATLTVDKFSRTAYMELSSLTSEDSAVYYCARSGAYSSDYSYDGFAYWGQGTLVTVSAASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKGLPAPIEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO: 9GATATTGTGATGACCCAGGCTGCATTCTCCAATCCAGTCACTCTTGGAACATCAGCTTCCATCTCCTGCAGGTCTAGTAAGAGTCTCCTACATAGTAATGGCATCACTTATTTGTATTGGTATCTGCAGAAGCCAGGCCAGTCTCCTCAGCTCCTGATTTATCAGATGTCCAACCTTGCCTCAGGAGTCCCAGACAGGTTCAGTGGCAGTGGGTCAGGAACTGCTTTCACACTGAGAATCAGTAGAGTGGAGGCTGAGGATGTGGGTGTTTATTACTGTATGCAACATCTAGAATATCCGTACACGTTCGGAGGGGGGACCAAGCTGGAAATAAAACGGGCTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAG SEQ ID NO: 10DIVMTQAAFSNPVTLGTSASISCRSSKSLLHSNGITYLYWYLQKPGQSPQLLIYQMSNLASGVPDRFSGSGSGTAFTLRISRVEAEDVGVYYCMQHLEYPYTFGGGTKLEIKVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQ ID NO: 11CAGGTCCAAGTGCAGCAGCCTGGGGCTGAAATTGTGAGGCCTGGGGCTTCAGTGAAGCTGTCCTGCAAGGCTTCTGGCTACACCTTCACCAGCTACTGGATGCACTGGGTGAAGCAGAGGCCTGGACAAGGCCTTGAGTGGATTGGACTGATTAATCCTACCAACGGTCGTACTAACTACAATGAGAAGTTCAAGAGCAAGGCCACACTGACTGTAGACAAATCCTCCAGCACAGCCTACATGCAACTCAGCAGCCTGACATCTGAGGACTCTGCGGTCTATTACTGTGCAAGAGGGGGGGACGGGGACTACTTTGACTACTGGGGCCAAGGCACCACTCTCACAGTCTCCTCAGCCTCAACGAAGGGCCCATCGGTCTTCCCCCTGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCTCTGACCAGCGGCGTGCACACCTTCCCAGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAACTTCGGCACCCAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGACAGTTGAGCGCAAATGTTGTGTCGAGTGCCCACCGTGCCCAGCACCACCTGTGGCAGGACCGTCAGTCTTCCGCTTCCCCCCAAAACCCAAGGACACCCGCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGATGTGAGCCACGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCACGGGAGGAGCAGTTCAACAGCACGTTCCGTGTGGTCAGCGTCCTCACCGTTGTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGGCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAACCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACACCTCCCATGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAATGA SEQ ID NO: 12QVQVQQPGAEIVRPGASVKLSCKASGYTFTSYWMHWVKQRPGQGLEWIGLINPTNGRTNYNEKFKSKATLTVDKSSSTAYMQLSSLTSEDSAVYYCARGGDGDYFDYWGQGTTLTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKGLPAPIEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO: 13GATATTGTGATGACCCAGGCTGCACCCTCTGTACCTGTCACTCCTGGAGAGTCAGTATCCATCTCCTGCAGGTCTACTAAGAGTCTCCTGCATAGTAATGGCAACACTTACTTGTATTGGTTCCTGCAGAGGCCAGGCCAGTCTCCTCAGCTCCTGATATATCGGATGTCCAACCTTGCCTCAGGAGTCCCAGACAGGTTCAGTGGCAGTGGGTCAGGAACTGCTTTCACACTGAGAATCAGTAGAGTGGAGGCTGAGGATGTGGGTGTTTATTACTGTATGCAACATCTAGAATATCCTTTCACGTTCGGAGGGGGGACCAAGCTGGAAATAAAACGGGCTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAG SEQ ID NO: 14DIVMTQAAPSVPVTPGESVSISCRSTKSLLHSNGNTYLYWFLQRPGQSPQLLIYRMSNLASGVPDRFSGSGSGTAFTLRISRVEAEDVGVYYCMQHLEYPFTFGGGTKLEIKVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQ ID NO: 15GAGATCCAGCTGCAGCAGTCTGGAGTTGAGCTGGTGAGGCCTGGGGCTTCAGTGACGCTGTCCTGCAAGGCTTCGGGCTACACATTTACTGACTATGACATGCACTGGGTGAAGCAGACACCTGTTCATGGCCTGGAATGGATTGGAACTATTGATCCTGAAACTGGTGGTACTGCCTACAATCAGAAGTTCAAGGGCAAGGCCACACTGACTGCGGACAGATCCTCCACCACAGCCTACATGGAGCTCAGCAGCCTGACATCTGAGGACTCTGCCGTCTATTACTGTACAACTTTCTACTATAGTCACTATAATTACGACGTGGGGTTTGCTTACTGGGGCCAAGGGACTCTGGTCACTGTCTCTGCAGCCTCAACGAAGGGCCCATCGGTCTTCCCCCTGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCTCTGACCAGCGGCGTGCACACCTTCCCAGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAACTTCGGCACCCAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGACAGTTGAGCGCAAATGTTGTGTCGAGTGCCCACCGTGCCCAGCACCACCTGTGGCAGGACCGTCAGTCTTCCGCTTCCCCCCAAAACCCAAGGACACCCGCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGATGTGAGCCACGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCACGGGAGGAGCAGTTCAACAGCACGTTCCGTGTGGTCAGCGTCCTCACCGTTGTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGGCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAACCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACACCTCCCATGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAATGA SEQ ID NO: 16EIQLQQSGVELVRPGASVTLSCKASGYTFTDYDMHWVKQTPVHGLEWIGTIDPETGGTAYNQKFKGKATLTADRSSTTAYMELSSLTSEDSAVYYCTTFYYSHYNYDVGFAYWGQGTLVTVSAASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKGLPAPIEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO: 17GATATTGTGATGACCCAGGCTGCACCCTCTGTACCTGTCACTCCTGGAGAGTCAGTATCCATCTCCTGCAGGTCTAGTAAGAGTCTCCTGCATAGTAATGGCAACACTTACTTGTATTGGTTCCTGCAGAGGCCAGGCCAGTCCCCTCAGCTCCTGATATATCGGATGTCCAACCTTGCCTCAGGAGTCCCAGACAGGTTCAGTGGCAGTGGGTCAGGAACTGCTTTCACACTGAGAATCAGTAGAGTGGAGGCTGAGGATGTGGGTGTTTATTACTGTATGCAACATCTAGAATATCCTTTCACGTTCGGAGGGGGGACCAAGCTGGAAATAAAACGGGCTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAG SEQ ID NO: 18DIVMTQAAPSVPVTPGESVSISCRSSKSLLHSNGNTYLYWFLQRPGQSPQLLIYRMSNLASGVPDRFSGSGSGTAFTLRISRVEAEDVGVYYCMQHLEYPFTFGGGTKLEIKVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQ ID NO: 19GAGATCCAGCTGCAGCAGTCTGGAGCTGAGCTGGTGAGGCCTGGGGCTTCAGTGACGCTGTCCTGCAAGGCTTCGGGCTACACATTTACTGACTATGAAATGCACTGGGTGAAGCAGACACCTGTTCATGGCCTGGAATGGATTGGAGCTATTGATCCTGAAACTGGTGGTACTGCCTACAATCAGAAGTTCAAGGGCAAGGCCACACTGACTGCAGACAAATCCTCCAGCACAGCCTACATGGAGCTCAGCAGCCTGACATCTGAGGACTCTGCCGTCTATTACTGTACAAGTTTCTACTATACTTACTATAATTACGACGTGGGGTTTGCTTACTGGGGCCAAGGGACTCTGGTCACTGTCTCTGCAGCCTCAACTGGGGCGTCTTATTACTATGCTATGGACCACTGGGGTCAAGGAACCTCAGTCACCGTCTCCTCAGCCTCAACGAAGGGCCCATCGGTCTTCCCCCTGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCTCTGACCAGCGGCGTGCACACCTTCCCAGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAACTTCGGCACCCAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGACAGTTGAGCGCAAATGTTGTGTCGAGTGCCCACCGTGCCCAGCACCACCTGTGGCAGGACCGTCAGTCTTCCGCTTCCCCCCAAAACCCAAGGACACCCGCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGATGTGAGCCACGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCACGGGAGGAGCAGTTCAACAGCACGTTCCGTGTGGTCAGCGTCCTCACCGTTGTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGGCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAACCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACACCTCCCATGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAATGA SEQ ID NO: 20EIQLQQSGAELVRPGASVTLSCKASGYTFTDYEMHWVKQTPVHGLEWIGAIDPETGGTAYNQKFKGKATLTADKSSSTAYMELSSLTSEDSAVYYCTSFYYTYYNYDVGFAYWGQGTLVTVSAASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKGLPAPIEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO: 21GATATTGTGATGACCCAGGCTGCATTCTCCAATCCAGTCACTCTTGGAACATCAGCTTCCATCTCCTGCAGGTCTAGTAAGAGTCTCCTACATAGTAATGGCATCACTTATTTGTATTGGTATCTGCAGAAGCCAGGCCAGTCTCCTCAGCTCCTGATTTATCAGATGTCCAACCTTGCCTCAGGAGTCCCAGACAGGTTCAGTAGCAGTGGGTCAGGAACTGATTTCACACTGAGAATCAGCAGAGTGGAGGCTGAGGATGTGGGTGTTTATTACTGTGCTCAAAATCTAGAACTTCCGTACACGTTCGGAGGGGGGACCAAGCTGGAAATAAAACGGGCTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAG SEQ ID NO: 22DIVMTQAAFSNPVTLGTSASISCRSSKSLLHSNGITYLYWYLQKPGQSPQLLIYQMSNLASGVPDRFSSSGSGTDFTLRISRVEAEDVGVYYCAQNLELPYTFGGGTKLEIKVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQ ID NO: 23CAGGTCCAAGTGCAGCAGCCTGGGGCTGAGCTTGTGAAGCCTGGGGCTTCGGTGAAGCTGTCCTGCAAGGCTTCTGGCTACACCTTCACCAGCTACTGGATGCACTGGGTGAAGCAGAGGCCTGGACAAGGCCTTGAGTGGATTGGACTGATTAATCCTAGCAACGCTCGTACTAACTACAATGAGAAGTTCAATACCAAGGCCACACTGACTGTAGACAAATCCTCCAGCACAGCCTACATGCAACTCAGCAGCCTGACATCTGAGGACTCTGCGGTCTATTACTGTGCAAGAGGGGGGGACGGGGACTACTTTGACTACTGGGGCCAAGGCACCACTCTCACAGTCTCCTCAGCCTCAACGAAGGGCCCATCGGTCTTCCCCCTGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCTCTGACCAGCGGCGTGCACACCTTCCCAGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAACTTCGGCACCCAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGACAGTTGAGCGCAAATGTTGTGTCGAGTGCCCACCGTGCCCAGCACCACCTGTGGCAGGACCGTCAGTCTTCCGCTTCCCCCCAAAACCCAAGGACACCCGCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGATGTGAGCCACGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCACGGGAGGAGCAGTTCAACAGCACGTTCCGTGTGGTCAGCGTCCTCACCGTTGTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGGCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAACCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACACCTCCCATGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAATGA SEQ ID NO: 24QVQVQQPGAELVKPGASVKLSCKASGYTFTSYWMHWVKQRPGQGLEWIGLINPSNARTNYNEKFNTKATLTVDKSSSTAYMQLSSLTSEDSAVYYCARGGDGDYFDYWGQGTTLTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKGLPAPIEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO: 25CAAATTGTTCTCACCCAGTCTCCAACACTCATGTCTGCATCTCCAGGGGAGAAGGTCACCATGACCTGCAGTGCCAGCTCAAGTGTAAGTTACATGTACTGGTACCAGCAGAAGCCAAGATCCTCCCCCAAACCCTGGATTTATCGCACATCCAACCTGGTTTCTGGAGTCCCTGTACGCTTCAGTGGCAGTGGGTCTGGGACCTCTTACTCTCTCACAATCAGCAGCATGGAGGCTGAAGATGCTGCCACTTATTACTGCCAGCAGTGGAGTAGTAACCCACCCACGTTCGGTGCTGGGACCAAGCTGGAGCTGAAACGGGCTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAG SEQ ID NO: 26QIVLTQSPTLMSASPGEKVTMTCSASSSVSYMYWYQQKPRSSPKPWIYRTSNLVSGVPVRFSGSGSGTSYSLTISSMEAEDAATYYCQQWSSNPPTFGAGTKLELKVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQ ID NO: 27GAAGTGAAGCTTGAGGAGTCTGGAGGTGGCCTGGTGCAGCCTGGAGGATCCCTGAAACTCTCCTGTGCAGCCTCAGGATTCGATTTTAGTAAAGACTGGATGAGTTGGGTCCGGCAGGCTCCAGGGAAAGGGCTAGAATGGATTGGAGAAATTAATCCAGATAGCAGTACGATAAACTATGCACCATCTCTTAAGGATAAATTCATCATCTCCAGAGAGAACGCCAAAAATACGCTGTACCTGCAAATGAGCAAAGTGAGATCTGAGGACACAGCCCTTTATTACTGTTCAAGACTAGAGGACTACGAAGACTGGTACTTCGATGTCTGGGGCGCAGGGACCACGGTCACCGTCTCCTCAGCCTCAACGAAGGGCCCATCGGTCTTCCCCCTGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCTCTGACCAGCGGCGTGCACACCTTCCCAGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAACTTCGGCACCCAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGACAGTTGAGCGCAAATGTTGTGTCGAGTGCCCACCGTGCCCAGCACCACCTGTGGCAGGACCGTCAGTCTTCCGCTTCCCCCCAAAACCCAAGGACACCCGCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGATGTGAGCCACGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCACGGGAGGAGCAGTTCAACAGCACGTTCCGTGTGGTCAGCGTCCTCACCGTTGTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGGCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAACCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACACCTCCCATGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAATGA SEQ ID NO: 28EVKLEESGGGLVQPGGSLKLSCAASGFDFSKDWMSWVRQAPGKGLEWIGEINPDSSTINYAPSLKDKFIISRENAKNTLYLQMSKVRSEDTALYYCSRLEDYEDWYFDVWGAGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKGLPAPIEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO: 29AGTATTGTGATGACCCAGACTCCCAAATTCCTGCTTGTATCAGCAGGAGACAGGGTTACCATAACCTGCAAGGCCAGTCAGAGTGTGAGTAATGCTGTAGCTTGGTACCAACAGAAGCCAGGGCAGTCTCCTAAACTGCTGATATACTATACATCCAATCGCTACACTGGAGTCCCTGATCGCTTCACTGGCAGTGGATATGGGACGGATTTCACTTTCACCATCACCACTGTGCAGGCTGAAGACCTGGCAGTTTATTTCTGTCAGCAGGATTATACCTCTCCGTGGACGTTCGGTGGAGGCACCAAGCTGGAAATCAAACGGGCTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAG SEQ ID NO: 30SIVMTQTPKFLLVSAGDRVTITCKASQSVSNAVAWYQQKPGQSPKLLIYYTSNRYTGVPDRFTGSGYGTDFTFTITTVQAEDLAVYFCQQDYTSPWTFGGGTKLEIKVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQ ID NO: 31CAGGTCCAACTGCAGCAGCCTGGGGCTGAACTGGCGAAGCCTGGGGCTTCAGTGAAGTTGTCCTGCAAGGCTTCTGGCTACACCTTCAACACCTATAATATGTACTGGTTGAAACAGAGGCCTGGGCAAGGCCTTGAGTGGATTGGGGGGATTGATCCTAGCAATGGTGATACTAAAATCAATGAGAAGTTCAAGAACAAGGCCACACTGACTGTTGACAAATCCTCCAGTACAGCCTATATGCAACTCAGCGGCCTGACATCTGAGGACTCTGCGGTCTATTACTGTACAAGCCATACGTACTGGGGCCAAGGGACTCTGGTCACTGTCTCTGCAGCCTCAACGAAGGGCCCATCGGTCTTCCCCCTGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCTCTGACCAGCGGCGTGCACACCTTCCCAGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAACTTCGGCACCCAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGACAGTTGAGCGCAAATGTTGTGTCGAGTGCCCACCGTGCCCAGCACCACCTGTGGCAGGACCGTCAGTCTTCCGCTTCCCCCCAAAACCCAAGGACACCCGCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGATGTGAGCCACGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCACGGGAGGAGCAGTTCAACAGCACGTTCCGTGTGGTCAGCGTCCTCACCGTTGTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGGCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAACCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACACCTCCCATGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAATGA SEQ ID NO: 32QVQLQQPGAELAKPGASVKLSCKASGYTFNTYNMYWLKQRPGQGLEWIGGIDPSNGDTKINEKFKNKATLTVDKSSSTAYMQLSGLTSEDSAVYYCTSHTYWGQGTLVTVSAASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKGLPAPIEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSL SPGKSEQ ID NO: 33GATATTGTGATGACCCAGGCTGCACCCTCTGTACCTGTCACTCCTGGAGAGTCAGTATCCATCTCCTGCAGGTCTACTAAGAGTCTCCTGCATAGTAATGGCAACACTTACTTGTATTGGTTCCTGCAGAGGCCAGGCCAGTCTCCTCAGCTCCTGATATATCGGATGTCCAACCTTGCCTCAGGAGTCCCAGACAGGTTCAGTGGCAGTGGGTCAGGAACTGCTTTCACACTGAGAATCAGTAGAGTGGAGGCTGAGGATGTGGGTGTTTATTACTGTATGCAACATCTAGAATATCCTTTCACGTTCGGAGGGGGGACCAAGCTGGAAATAAAACGGGCTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAG SEQ ID NO: 34DIVMTQAAPSVPVTPGESVSISCRSTKSLLHSNGNTYLYWFLQRPGQSPQLLIYRMSNLASGVPDRFSGSGSGTAFTLRISRVEAEDVGVYYCMQHLEYPFTFGGGTKLEIKVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQ ID NO: 35GAGATCCAGCTGCAGCAGTCTGGAGTTGAGCTGGTGAGGCCTGGGGCTTCAGTGACGCTGTCCTGCAAGGCTTCGGGCTACACATTTACTGACTATGACATGCACTGGGTGAAGCAGACACCTGTTCATGGCCTGGAATGGATTGGAACTATTGATCCTGAAACTGGTGGTACTGCCTACAATCAGAAGTTCAAGGGCAAGGCCACACTGACTGCGGACAGATCCTCCACCACAGCCTACATGGAGCTCAGCAGCCTGACATCTGAGGACTCTGCCGTCTATTACTGTACAAGTTTCTACTATACTTACTCTAATTACGACGTGGGGTTTGCTTACTGGGGCCAAGGGACTCTGGTCACTGTCTCTGCAGCCTCAACTGGGGCGTCTTATTACTATGCTATGGACCACTGGGGTCAAGGAACCTCAGTCACCGTCTCCTCAGCCTCAACGAAGGGCCCATCGGTCTTCCCCCTGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCTCTGACCAGCGGCGTGCACACCTTCCCAGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAACTTCGGCACCCAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGACAGTTGAGCGCAAATGTTGTGTCGAGTGCCCACCGTGCCCAGCACCACCTGTGGCAGGACCGTCAGTCTTCCGCTTCCCCCCAAAACCCAAGGACACCCGCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGATGTGAGCCACGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCACGGGAGGAGCAGTTCAACAGCACGTTCCGTGTGGTCAGCGTCCTCACCGTTGTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGGCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAACCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACACCTCCCATGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAATGA SEQ ID NO: 36EIQLQQSGVELVRPGASVTLSCKASGYTFTDYDMHWVKQTPVHGLEWIGTIDPETGGTAYNQKFKGKATLTADRSSTTAYMELSSLTSEDSAVYYCTSFYYTYSNYDVGFAYWGQGTLVTVSAASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKGLPAPIEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO: 37GAAAATGTGCTCACCCAGTCTCCAGCAATCATGTCTGCATCTCCAGGGGAGAAGGTCACCATATCCTGCAGTGCCAGCTCAAGTGTAAGTTACATGTACTGGTACCAGCAGAAGCCAGGATCCTCCCCCAAACCCTGGATTTATCGCACATCCAACCTGGCTTCTGGAGTCCCTGCTCGCTTCAGTGGCAGTGGGTCTGGGACCTCTTACTCTCTCACAATCAGCAGCATGGAGGCTGAAGATGCTGCCACTTATTACTGCCAGCAGTGGAGTAGTAACCCACTCACGTTCGGTGCTGGGACCAAGCTGG AGCTGAAASEQ ID NO: 38ENVLTQSPAIMSASPGEKVTISCSASSSVSYMYWYQQKPGSSPKPWIYRTSNLASGVPARFSGSGSGTSYSLTISSMEAEDAATYYCQQWSSNPLTFGAGTKLELK SEQ ID NO: 39GAGGTCCAGCTGCAACAATCTGGGACTGAGCTGGTGAGGCCTGGGTCCTCAGTGAAGATTTCCTGCAAGGCTTCTGGCTACACCTTCACCAGGTACTGGATGGACTGGGTGAAGCAGAGGCCTGGACAAGGCCTTGAGTGGATCGGAGAGATTGATCCTTCTGATAGTTATACTAACTACAATCAAAAGTTCAAGGGCAAGGCCACATTGACTGTAGATAAATTCTCCAGAACAGCCTATATGGAACTCAGCAGCCTGACATCTGAGGACTCTGCGGTCTATTACTGTGCAAGATCGGGGGCCTACTCTAGTGACTATAGTTACGACGGGTTTGCTTACTGGGGCCAAGGGACTCTGGTCACTGTCTCTGCASEQ ID NO: 40EVQLQQSGTELVRPGSSVKISCKASGYTFTRYWMDWVKQRPGQGLEWIGEIDPSDSYTNYNQKFKGKATLTVDKFSRTAYMELSSLTSEDSAVYYCARSGAYSSDYSYDGFAYWGQGTLVTVSASEQ ID NO: 41GATATTGTGATGACCCAGGCTGCATTCTCCAATCCAGTCACTCTTGGAACATCAGCTTCCATCTCCTGCAGGTCTAGTAAGAGTCTCCTACATAGTAATGGCATCACTTATTTGTATTGGTATCTGCAGAAGCCAGGCCAGTCTCCTCAGCTCCTGATTTATCAGATGTCCAACCTTGCCTCAGGAGTCCCAGACAGGTTCAGTGGCAGTGGGTCAGGAACTGCTTTCACACTGAGAATCAGTAGAGTGGAGGCTGAGGATGTGGGTGTTTATTACTGTATGCAACATCTAGAATATCCGTACACGTTCGGAGGGGGGACCAAGCTGGAAATAAAA SEQ ID NO: 42DIVMTQAAFSNPVTLGTSASISCRSSKSLLHSNGITYLYWYLQKPGQSPQLLIYQMSNLASGVPDRFSGSGSGTAFTLRISRVEAEDVGVYYCMQHLEYPYTFGGGTKLEIK SEQ ID NO: 43CAGGTCCAAGTGCAGCAGCCTGGGGCTGAAATTGTGAGGCCTGGGGCTTCAGTGAAGCTGTCCTGCAAGGCTTCTGGCTACACCTTCACCAGCTACTGGATGCACTGGGTGAAGCAGAGGCCTGGACAAGGCCTTGAGTGGATTGGACTGATTAATCCTACCAACGGTCGTACTAACTACAATGAGAAGTTCAAGAGCAAGGCCACACTGACTGTAGACAAATCCTCCAGCACAGCCTACATGCAACTCAGCAGCCTGACATCTGAGGACTCTGCGGTCTATTACTGTGCAAGAGGGGGGGACGGGGACTACTTTGACTACTGGGGCCAAGGCACCACTCTCACAGTCTCCTCA SEQ ID NO: 44QVQVQQPGAEIVRPGASVKLSCKASGYTFTSYWMHWVKQRPGQGLEWIGLINPTNGRTNYNEKFKSKATLTVDKSSSTAYMQLSSLTSEDSAVYYCARGGDGDYFDYWGQGTTLTVSS SEQ ID NO: 45GATATTGTGATGACCCAGGCTGCACCCTCTGTACCTGTCACTCCTGGAGAGTCAGTATCCATCTCCTGCAGGTCTACTAAGAGTCTCCTGCATAGTAATGGCAACACTTACTTGTATTGGTTCCTGCAGAGGCCAGGCCAGTCTCCTCAGCTCCTGATATATCGGATGTCCAACCTTGCCTCAGGAGTCCCAGACAGGTTCAGTGGCAGTGGGTCAGGAACTGCTTTCACACTGAGAATCAGTAGAGTGGAGGCTGAGGATGTGGGTGTTTATTACTGTATGCAACATCTAGAATATCCTTTCACGTTCGGAGGGGGGACCAAGCTGGAAATAAAA SEQ ID NO: 46DIVMTQAAPSVPVTPGESVSISCRSTKSLLHSNGNTYLYWFLQRPGQSPQLLIYRMSNLASGVPDRFSGSGSGTAFTLRISRVEAEDVGVYYCMQHLEYPFTFGGGTKLEIK SEQ ID NO: 47GAGATCCAGCTGCAGCAGTCTGGAGTTGAGCTGGTGAGGCCTGGGGCTTCAGTGACGCTGTCCTGCAAGGCTTCGGGCTACACATTTACTGACTATGACATGCACTGGGTGAAGCAGACACCTGTTCATGGCCTGGAATGGATTGGAACTATTGATCCTGAAACTGGTGGTACTGCCTACAATCAGAAGTTCAAGGGCAAGGCCACACTGACTGCGGACAGATCCTCCACCACAGCCTACATGGAGCTCAGCAGCCTGACATCTGAGGACTCTGCCGTCTATTACTGTACAACTTTCTACTATAGTCACTATAATTACGACGTGGGGTTTGCTTACTGGGGCCAAGGGACTCTGGTCACTGTCTCTGCASEQ ID NO: 48EIQLQQSGVELVRPGASVTLSCKASGYTFTDYDMHWVKQTPVHGLEWIGTIDPETGGTAYNQKFKGKATLTADRSSTTAYMELSSLTSEDSAVYYCTTFYYSHYNYDVGFAYWGQGTLVTVSASEQ ID NO: 49GATATTGTGATGACCCAGGCTGCACCCTCTGTACCTGTCACTCCTGGAGAGTCAGTATCCATCTCCTGCAGGTCTAGTAAGAGTCTCCTGCATAGTAATGGCAACACTTACTTGTATTGGTTCCTGCAGAGGCCAGGCCAGTCCCCTCAGCTCCTGATATATCGGATGTCCAACCTTGCCTCAGGAGTCCCAGACAGGTTCAGTGGCAGTGGGTCAGGAACTGCTTTCACACTGAGAATCAGTAGAGTGGAGGCTGAGGATGTGGGTGTTTATTACTGTATGCAACATCTAGAATATCCTTTCACGTTCGGAGGGGGGACCAAGCTGGAAATAAAA SEQ ID NO: 50DIVMTQAAPSVPVTPGESVSISCRSSKSLLHSNGNTYLYWFLQRPGQSPQLLIYRMSNLASGVPDRFSGSGSGTAFTLRISRVEAEDVGVYYCMQHLEYPFTFGGGTKLEIK SEQ ID NO: 51GAGATCCAGCTGCAGCAGTCTGGAGCTGAGCTGGTGAGGCCTGGGGCTTCAGTGACGCTGTCCTGCAAGGCTTCGGGCTACACATTTACTGACTATGAAATGCACTGGGTGAAGCAGACACCTGTTCATGGCCTGGAATGGATTGGAGCTATTGATCCTGAAACTGGTGGTACTGCCTACAATCAGAAGTTCAAGGGCAAGGCCACACTGACTGCAGACAAATCCTCCAGCACAGCCTACATGGAGCTCAGCAGCCTGACATCTGAGGACTCTGCCGTCTATTACTGTACAAGTTTCTACTATACTTACTATAATTACGACGTGGGGTTTGCTTACTGGGGCCAAGGGACTCTGGTCACTGTCTCTGCASEQ ID NO: 52EIQLQQSGAELVRPGASVTLSCKASGYTFTDYEMHWVKQTPVHGLEWIGAIDPETGGTAYNQKFKGKATLTADKSSSTAYMELSSLTSEDSAVYYCTSFYYTYYNYDVGFAYWGQGTLVTVSASEQ ID NO: 53GATATTGTGATGACCCAGGCTGCATTCTCCAATCCAGTCACTCTTGGAACATCAGCTTCCATCTCCTGCAGGTCTAGTAAGAGTCTCCTACATAGTAATGGCATCACTTATTTGTATTGGTATCTGCAGAAGCCAGGCCAGTCTCCTCAGCTCCTGATTTATCAGATGTCCAACCTTGCCTCAGGAGTCCCAGACAGGTTCAGTAGCAGTGGGTCAGGAACTGATTTCACACTGAGAATCAGCAGAGTGGAGGCTGAGGATGTGGGTGTTTATTACTGTGCTCAAAATCTAGAACTTCCGTACACGTTCGGAGGGGGGACCAAGCTGGAAATAAAA SEQ ID NO: 54DIVMTQAAFSNPVTLGTSASISCRSSKSLLHSNGITYLYWYLQKPGQSPQLLIYQMSNLASGVPDRFSSSGSGTDFTLRISRVEAEDVGVYYCAQNLELPYTFGGGTKLEIK SEQ ID NO: 55CAGGTCCAAGTGCAGCAGCCTGGGGCTGAGCTTGTGAAGCCTGGGGCTTCGGTGAAGCTGTCCTGCAAGGCTTCTGGCTACACCTTCACCAGCTACTGGATGCACTGGGTGAAGCAGAGGCCTGGACAAGGCCTTGAGTGGATTGGACTGATTAATCCTAGCAACGCTCGTACTAACTACAATGAGAAGTTCAATACCAAGGCCACACTGACTGTAGACAAATCCTCCAGCACAGCCTACATGCAACTCAGCAGCCTGACATCTGAGGACTCTGCGGTCTATTACTGTGCAAGAGGGGGGGACGGGGACTACTTTGACTACTGGGGCCAAGGCACCACTCTCACAGTCTCCTCA SEQ ID NO: 56QVQVQQPGAELVKPGASVKLSCKASGYTFTSYWMHWVKQRPGQGLEWIGLINPSNARTNYNEKFNTKATLTVDKSSSTAYMQLSSLTSEDSAVYYCARGGDGDYFDYWGQGTTLTVSS SEQ ID NO: 57CAAATTGTTCTCACCCAGTCTCCAACACTCATGTCTGCATCTCCAGGGGAGAAGGTCACCATGACCTGCAGTGCCAGCTCAAGTGTAAGTTACATGTACTGGTACCAGCAGAAGCCAAGATCCTCCCCCAAACCCTGGATTTATCGCACATCCAACCTGGTTTCTGGAGTCCCTGTACGCTTCAGTGGCAGTGGGTCTGGGACCTCTTACTCTCTCACAATCAGCAGCATGGAGGCTGAAGATGCTGCCACTTATTACTGCCAGCAGTGGAGTAGTAACCCACCCACGTTCGGTGCTGGGACCAAGCTGG AGCTGAAASEQ ID NO: 58QIVLTQSPTLMSASPGEKVTMTCSASSSVSYMYWYQQKPRSSPKPWIYRTSNLVSGVPVRFSGSGSGTSYSLTISSMEAEDAATYYCQQWSSNPPTFGAGTKLELK SEQ ID NO: 59GAAGTGAAGCTTGAGGAGTCTGGAGGTGGCCTGGTGCAGCCTGGAGGATCCCTGAAACTCTCCTGTGCAGCCTCAGGATTCGATTTTAGTAAAGACTGGATGAGTTGGGTCCGGCAGGCTCCAGGGAAAGGGCTAGAATGGATTGGAGAAATTAATCCAGATAGCAGTACGATAAACTATGCACCATCTCTTAAGGATAAATTCATCATCTCCAGAGAGAACGCCAAAAATACGCTGTACCTGCAAATGAGCAAAGTGAGATCTGAGGACACAGCCCTTTATTACTGTTCAAGACTAGAGGACTACGAAGACTGGTACTTCGATGTCTGGGGCGCAGGGACCACGGTCACCGTCTCCTCA SEQ ID NO: 60EVKLEESGGGLVQPGGSLKLSCAASGFDFSKDWMSWVRQAPGKGLEWIGEINPDSSTINYAPSLKDKFIISRENAKNTLYLQMSKVRSEDTALYYCSRLEDYEDWYFDVWGAGTTVTVSS SEQ ID NO: 61AGTATTGTGATGACCCAGACTCCCAAATTCCTGCTTGTATCAGCAGGAGACAGGGTTACCATAACCTGCAAGGCCAGTCAGAGTGTGAGTAATGCTGTAGCTTGGTACCAACAGAAGCCAGGGCAGTCTCCTAAACTGCTGATATACTATACATCCAATCGCTACACTGGAGTCCCTGATCGCTTCACTGGCAGTGGATATGGGACGGATTTCACTTTCACCATCACCACTGTGCAGGCTGAAGACCTGGCAGTTTATTTCTGTCAGCAGGATTATACCTCTCCGTGGACGTTCGGTGGAGGCACCAAGCTGGAAATCAAA SEQ ID NO: 62SIVMTQTPKFLLVSAGDRVTITCKASQSVSNAVAWYQQKPGQSPKLLIYYTSNRYTGVPDRFTGSGYGTDFTFTITTVQAEDLAVYFCQQDYTSPWTFGGGTKLEIK SEQ ID NO: 63CAGGTCCAACTGCAGCAGCCTGGGGCTGAACTGGCGAAGCCTGGGGCTTCAGTGAAGTTGTCCTGCAAGGCTTCTGGCTACACCTTCAACACCTATAATATGTACTGGTTGAAACAGAGGCCTGGGCAAGGCCTTGAGTGGATTGGGGGGATTGATCCTAGCAATGGTGATACTAAAATCAATGAGAAGTTCAAGAACAAGGCCACACTGACTGTTGACAAATCCTCCAGTACAGCCTATATGCAACTCAGCGGCCTGACATCTGAGGACTCTGCGGTCTATTACTGTACAAGCCATACGTACTGGGGCCAAGGGACTCTGGTCACTGTCTCTGCA SEQ ID NO: 64QVQLQQPGAELAKPGASVKLSCKASGYTFNTYNMYWLKQRPGQGLEWIGGIDPSNGDTKINEKFKNKATLTVDKSSSTAYMQLSGLTSEDSAVYYCTSHTYWGQGTLVTVSA SEQ ID NO: 65GATATTGTGATGACCCAGGCTGCACCCTCTGTACCTGTCACTCCTGGAGAGTCAGTATCCATCTCCTGCAGGTCTACTAAGAGTCTCCTGCATAGTAATGGCAACACTTACTTGTATTGGTTCCTGCAGAGGCCAGGCCAGTCTCCTCAGCTCCTGATATATCGGATGTCCAACCTTGCCTCAGGAGTCCCAGACAGGTTCAGTGGCAGTGGGTCAGGAACTGCTTTCACACTGAGAATCAGTAGAGTGGAGGCTGAGGATGTGGGTGTTTATTACTGTATGCAACATCTAGAATATCCTTTCACGTTCGGAGGGGGGACCAAGCTGGAAATAAAA SEQ ID NO: 66DIVMTQAAPSVPVTPGESVSISCRSTKSLLHSNGNTYLYWFLQRPGQSPQLLIYRMSNLASGVPDRFSGSGSGTAFTLRISRVEAEDVGVYYCMQHLEYPFTFGGGTKLEIK SEQ ID NO: 67GAGATCCAGCTGCAGCAGTCTGGAGTTGAGCTGGTGAGGCCTGGGGCTTCAGTGACGCTGTCCTGCAAGGCTTCGGGCTACACATTTACTGACTATGACATGCACTGGGTGAAGCAGACACCTGTTCATGGCCTGGAATGGATTGGAACTATTGATCCTGAAACTGGTGGTACTGCCTACAATCAGAAGTTCAAGGGCAAGGCCACACTGACTGCGGACAGATCCTCCACCACAGCCTACATGGAGCTCAGCAGCCTGACATCTGAGGACTCTGCCGTCTATTACTGTACAAGTTTCTACTATACTTACTCTAATTACGACGTGGGGTTTGCTTACTGGGGCCAAGGGACTCTGGTCACTGTCTCTGCASEQ ID NO: 68EIQLQQSGVELVRPGASVTLSCKASGYTFTDYDMHWVKQTPVHGLEWIGTIDPETGGTAYNQKFKGKATLTADRSSTTAYMELSSLTSEDSAVYYCTSFYYTYSNYDVGFAYWGQGTLVTVSASEQ ID NO: 69 SASSSVSYMY SEQ ID NO: 70 RTSNLAS SEQ ID NO: 71 QQWSSNPLTSEQ ID NO: 72 GYTFTRYWMD SEQ ID NO: 73 EIDPSDSYTN SEQ ID NO: 74ARSGAYSSDYSYDGFAY SEQ ID NO: 75 RSSKSLLHSNGITYLY SEQ ID NO: 76 QMSNLASSEQ ID NO: 77 MQHLEYPYT SEQ ID NO: 78 GYTFTSYWMH SEQ ID NO: 79LINPTNGRTN SEQ ID NO: 80 ARGGDGDYFDY SEQ ID NO: 81 RSTKSLLHSNGNTYLYSEQ ID NO: 82 RMSNLAS SEQ ID NO: 83 MQHLEYPFT SEQ ID NO: 84 GYTFTDYDMHSEQ ID NO: 85 TIDPETGGTA SEQ ID NO: 86 TTFYYSHYNYDVGFAY SEQ ID NO: 87RSSKSLLHSNGNTYLY SEQ ID NO: 88 RMSNLAS SEQ ID NO: 89 MQHLEYPFTSEQ ID NO: 90 GYTFTDYDMH SEQ ID NO: 91 AIDPETGGTA SEQ ID NO: 92TSFYYTYYNYDVGFAY SEQ ID NO: 93 RSSKSLLHSNGITYLY SEQ ID NO: 94 QMSNLASSEQ ID NO: 95 AQNLELPYT SEQ ID NO: 96 GYTFTSYWMH SEQ ID NO: 97LINPSNARTN SEQ ID NO: 98 ARGGDGDYFDY SEQ ID NO: 99 SASSSVSYMYSEQ ID NO: 100 RTSNLVS SEQ ID NO: 101 QQWSSNPPT SEQ ID NO: 102GFDFSKDWMS SEQ ID NO: 103 EINPDSSTIN SEQ ID NO: 104 SRLEDYEDWYFDVSEQ ID NO: 105 KASQSVSNAVA SEQ ID NO: 106 YTSNRYT SEQ ID NO: 107QQDYTSPWT SEQ ID NO: 108 GYTFNTYNMY SEQ ID NO: 109 GIDPSNGDTKSEQ ID NO: 110 TSHTY SEQ ID NO: 111 RSTKSLLHSNGNTYLY SEQ ID NO: 112RMSNLAS SEQ ID NO: 113 MQHLEYPFT SEQ ID NO: 114 GYTFTDYDMHSEQ ID NO: 115 TIDPETGGTA SEQ ID NO: 116 TSFYYTYSNYDVGFAY SEQ ID NO: 117GTAAGCAAGCTTGCTCACGCCTTCCGCGCGCTC SEQ ID NO: 118GTAAGCAGATCTCTGGCGCCATGGAAGCGGAACAG SEQ ID NO: 119CACTGGGAGCTATGGAAGAAGAC SEQ ID NO: 120 CAAAAGTGCAAAGAAGGGAAGACASEQ ID NO: 121 TGAAGGTCGGAGTCAACGGATTTGGT SEQ ID NO: 122CATGTGGGCCATGAGGTCCACCAC SEQ ID NO: 123VRTKIDTTENLLNTEVHSSPAQRWSMQVPPEVSAEAGDAAVLPCTFTHPHRHYDGPLTAIWRAGEPYAGPQVFRCAAARGSELCQTALSLHGRFRLLGNPRRNDLSLRVERLALADDRRYFCRVEFAGDVHDRYESRHGVRLHVTAAPRIVNISVLPSPAHAFRALCTAEGEPPPALAWSGPALGNSLAAVRSPREGHGHLVTAELPALTHDGRYTCTAANSLGRSEASVYLFRFHGASGAS SEQ ID NO: 124GTAAGCGGATCCGTGAGAACTAAAATAGATACTA SEQ ID NO: 125:GTAAGCGCGGCCGCGCTGGCGCCATGGAAGCGGAACAGGTA SEQ ID NO: 126GTACATTTATATTGGCTCATGTCCAATATGACCGCCATGTTGACATTGATTATTGACTAGTTATTAATAGTAATCAATTACGGGGTCATTAGTTCATAGCCCATATATGGAGTTCCGCGTTACATAACTTACGGTAAATGGCCCGCCTGGCTGACCGCCCAACGACCCCCGCCCATTGACGTCAATAATGACGTATGTTCCCATAGTAACGCCAATAGGGACTTTCCATTGACGTCAATGGGTGGAGTATTTACGGTAAACTGCCCACTTGGCAGTACATCAAGTGTATCATATGCCAAGTCCGCCCCCTATTGACGTCAATGACGGTAAATGGCCCGCCTGGCATTATGCCCAGTACATGACCTTACGGGACTTTCCTACTTGGCAGTACATCTACGTATTAGTCATCGCTATTACCATGGTGATGCGGTTTTGGCAGTACACCAATGGGCGTGGATAGCGGTTTGACTCACGGGGATTTCCAAGTCTCCACCCCATTGACGTCAATGGGAGTTTGTTTTGGCACCAAAATCAACGGGACTTTCCAAAATGTCGTAATAACCCCGCCCCGTTGACGCAAATGGGCGGTAGGCGTGTACGGTGGGAGGTCTATATAAGCAGAGCTCGTTTAGTGAACCGTCAGATCCTCACTCTCTTCCGCATCGCTGTCTGCGAGGGCCAGCTGTTGGGCTCGCGGTTGAGGACAAACTCTTCGCGGTCTTTCCAGTACTCTTGGATCGGAAACCCGTCGGCCTCCGAACGGTACTCCGCCACCGAGGGACCTGAGCCAGTCCGCATCGACCGGATCGGAAAACCTCTCGAGAAAGGCGTCTAACCAGTCACAGTCGCAAGGTAGGCTGAGCACCGTGGCGGGCGGCAGCGGGTGGCGGTCGGGGTTGTTTCTGGCGGAGGTGCTGCTGATGATGTAATTAAAGTAGGCGGTCTTGAGCCGGCGGATGGTCGAGGTGAGGTGTGGCAGGCTTGAGATCCAGCTGTTGGGGTGAGTACTCCCTCTCAAAAGCGGGCATGACTTCTGCGCTAAGATTGTCAGTTTCCAAAAACGAGGAGGATTTGATATTCACCTGGCCCGATCTGGCCATACACTTGAGTGACAATGACATCCACTTTGCCTTTCTCTCCACAGGTGTCCACTCCCAGGTCCAAGTTTGCCGCCACCATGGAGACAGACACACTCCTGCTATGGGTACTGCTGCTCTGGGTTCCAGGTTCCACTGGCGCCGGATCAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGTTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCCGGGAAAGCTAGCGGAGCCGGAAGCACAACCGAAAACCTGTATTTTCAGGGCGGATCCGAATTCAAGCTTGATATCTGATCCCCCGACCTCGACCTCTGGCTAATAAAGGAAATTTATTTTCATTGCAATAGTGTGTTGGAATTTTTTGTGTCTCTCACTCGGAAGGACATATGGGAGGGCAAATCATTTGGTCGAGATCCCTCGGAGATCTCTAGCTAGAGCCCCGCCGCCGGACGAACTAAACCTGACTACGGCATCTCTGCCCCTTCTTCGCGGGGCAGTGCATGTAATCCCTTCAGTTGGTTGGTACAACTTGCCAACTGAACCCTAAACGGGTAGCATATGCTTCCCGGGTAGTAGTATATACTATCCAGACTAACCCTAATTCAATAGCATATGTTACCCAACGGGAAGCATATGCTATCGAATTAGGGTTAGTAAAAGGGTCCTAAGGAACAGCGATGTAGGTGGGCGGGCCAAGATAGGGGCGCGATTGCTGCGATCTGGAGGACAAATTACACACACTTGCGCCTGAGCGCCAAGCACAGGGTTGTTGGTCCTCATATTCACGAGGTCGCTGAGAGCACGGTGGGCTAATGTTGCCATGGGTAGCATATACTACCCAAATATCTGGATAGCATATGCTATCCTAATCTATATCTGGGTAGCATAGGCTATCCTAATCTATATCTGGGTAGCATATGCTATCCTAATCTATATCTGGGTAGTATATGCTATCCTAATTTATATCTGGGTAGCATAGGCTATCCTAATCTATATCTGGGTAGCATATGCTATCCTAATCTATATCTGGGTAGTATATGCTATCCTAATCTGTATCCGGGTAGCATATGCTATCCTAATAGAGATTAGGGTAGTATATGCTATCCTAATTTATATCTGGGTAGCATATACTACCCAAATATCTGGATAGCATATGCTATCCTAATCTATATCTGGGTAGCATATGCTATCCTAATCTATATCTGGGTAGCATAGGCTATCCTAATCTATATCTGGGTAGCATATGCTATCCTAATCTATATCTGGGTAGTATATGCTATCCTAATTTATATCTGGGTAGCATAGGCTATCCTAATCTATATCTGGGTAGCATATGCTATCCTAATCTATATCTGGGTAGTATATGCTATCCTAATCTGTATCCGGGTAGCATATGCTATCCTCACGATGATAAGCTGTCAAACATGAGAATTAATTCTTGAAGACGAAAGGGCCTCGTGATACGCCTATTTTTATAGGTTAATGTCATGATAATAATGGTTTCTTAGACGTCAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGTATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTGTTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGCAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATTTAAAACTTCATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATCCCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCATTGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGTTCTTTCCTGCGTTATCCCCTGATTCTGTGGATAACCGTATTACCGCCTTTGAGTGAGCTGATACCGCTCGCCGCAGCCGAACGACCGAGCGCAGCGAGTCAGTGAGCGAGGAAGC SEQ ID NO: 127GTAAGCGCTAGCGCCTCAACGAAGGGCCCATCTGTCTTTCCCCTGGCCCC SEQ ID NO: 128GTAAGCGAATTCACAAGATTTGGGCTCAACTTTCTTG SEQ ID NO: 129GCTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAG SEQ ID NO: 130AVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQ ID NO: 131CTTGAGCCGGCGGATGGTCGAGGTGAGGTGTGGCAGGCTTGAGATCCAGCTGTTGGGGTGAGTACTCCCTCTCAAAAGCGGGCATTACTTCTGCGCTAAGATTGTCAGTTTCCAAAAACGAGGAGGATTTGATATTCACCTGGCCCGATCTGGCCATACACTTGAGTGACAATGACATCCACTTTGCCTTTCTCTCCACAGGTGTCCACTCCCAGGTCCAAGTTTAAACGGATCTCTAGCGAATTCATGAACTTTCTGCTGTCTTGGGTGCATTGGAGCCTTGCCTTGCTGCTCTACCTCCACCATGCCAAGTGGTCCCAGGCTTGAGACGGAGCTTACAGCGCTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAGGGTACCGCGGCCGCTTCGAATGAGATCCCCCGACCTCGACCTCTGGCTAATAAAGGAAATTTATTTTCATTGCAATAGTGTGTTGGAATTTTTTGTGTCTCTCACTCGGAAGGACATATGGGAGGGCAAATCATTTGGTCGAGATCCCTCGGAGATCTCTAGCTAGAGCCCCGCCGCCGGACGAACTAAACCTGACTACGGCATCTCTGCCCCTTCTTCGCGGGGCAGTGCATGTAATCCCTTCAGTTGGTTGGTACAACTTGCCAACTGGGCCCTGTTCCACATGTGACACGGGGGGGGACCAAACACAAAGGGGTTCTCTGACTGTAGTTGACATCCTTATAAATGGATGTGCACATTTGCCAACACTGAGTGGCTTTCATCCTGGAGCAGACTTTGCAGTCTGTGGACTGCAACACAACATTGCCTTTATGTGTAACTCTTGGCTGAAGCTCTTACACCAATGCTGGGGGACATGTACCTCCCAGGGGCCCAGGAAGACTACGGGAGGCTACACCAACGTCAATCAGAGGGGCCTGTGTAGCTACCGATAAGCGGACCCTCAAGAGGGCATTAGCAATAGTGTTTATAAGGCCCCCTTGTTAACCCTAAACGGGTAGCATATGCTTCCCGGGTAGTAGTATATACTATCCAGACTAACCCTAATTCAATAGCATATGTTACCCAACGGGAAGCATATGCTATCGAATTAGGGTTAGTAAAAGGGTCCTAAGGAACAGCGATATCTCCCACCCCATGAGCTGTCACGGTTTTATTTACATGGGGTCAGGATTCCACGAGGGTAGTGAACCATTTTAGTCACAAGGGCAGTGGCTGAAGATCAAGGAGCGGGCAGTGAACTCTCCTGAATCTTCGCCTGCTTCTTCATTCTCCTTCGTTTAGCTAATAGAATAACTGCTGAGTTGTGAACAGTAAGGTGTATGTGAGGTGCTCGAAAACAAGGTTTCAGGTGACGCCCCCAGAATAAAATTTGGACGGGGGGTTCAGTGGTGGCATTGTGCTATGACACCAATATAACCCTCACAAACCCCTTGGGCAATAAATACTAGTGTAGGAATGAAACATTCTGAATATCTTTAACAATAGAAATCCATGGGGTGGGGACAAGCCGTAAAGACTGGATGTCCATCTCACACGAATTTATGGCTATGGGCAACACATAATCCTAGTGCAATATGATACTGGGGTTATTAAGATGTGTCCCAGGCAGGGACCAAGACAGGTGAACCATGTTGTTACACTCTATTTGTAACAAGGGGAAAGAGAGTGGACGCCGACAGCAGCGGACTCCACTGGTTGTCTCTAACACCCCCGAAAATTAAACGGGGCTCCACGCCAATGGGGCCCATAAACAAAGACAAGTGGCCACTCTTTTTTTTGAAATTGTGGAGTGGGGGCACGCGTCAGCCCCCACACGCCGCCCTGCGGTTTTGGACTGTAAAATAAGGGTGTAATAACTTGGCTGATTGTAACCCCGCTAACCACTGCGGTCAAACCACTTGCCCACAAAACCACTAATGGCACCCCGGGGAATACCTGCATAAGTAGGTGGGCGGGCCAAGATAGGGGCGCGATTGCTGCGATCTGGAGGACAAATTACACACACTTGCGCCTGAGCGCCAAGCACAGGGTTGTTGGTCCTCATATTCACGAGGTCGCTGAGAGCACGGTGGGCTAATGTTGCCATGGGTAGCATATACTACCCAAATATCTGGATAGCATATGCTATCCTAATCTATATCTGGGTAGCATAGGCTATCCTAATCTATATCTGGGTAGCATATGCTATCCTAATCTATATCTGGGTAGTATATGCTATCCTAATTTATATCTGGGTAGCATAGGCTATCCTAATCTATATCTGGGTAGCATATGCTATCCTAATCTATATCTGGGTAGTATATGCTATCCTAATCTGTATCCGGGTAGCATATGCTATCCTAATAGAGATTAGGGTAGTATATGCTATCCTAATTTATATCTGGGTAGCATATACTACCCAAATATCTGGATAGCATATGCTATCCTAATCTATATCTGGGTAGCATATGCTATCCTAATCTATATCTGGGTAGCATAGGCTATCCTAATCTATATCTGGGTAGCATATGCTATCCTAATCTATATCTGGGTAGTATATGCTATCCTAATTTATATCTGGGTAGCATAGGCTATCCTAATCTATATCTGGGTAGCATATGCTATCCTAATCTATATCTGGGTAGTATATGCTATCCTAATCTGTATCCGGGTAGCATATGCTATCCTCACGATGATAAGCTGTCAAACATGAGAATTAATTCTTGAAGACGAAAGGGCCTCGTGATACGCCTATTTTTATAGGTTAATGTCATGATAATAATGGTTTCTTAGACGTCAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGTATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTGTTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGCAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATTTAAAACTTCATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATCCCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCATTGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGTTCTTTCCTGCGTTATCCCCTGATTCTGTGGATAACCGTATTACCGCCTTTGAGTGAGCTGATACCGCTCGCCGCAGCCGAACGACCGAGCGCAGCGAGTCAGTGAGCGAGGAAGCGGAAGAGCGCCCAATACGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGGCACGACAGGTTTCCCGACTGGAAAGCGGGCAGTGAGCGCAACGCAATTAATGTGAGTTAGCTCACTCATTAGGCACCCCAGGCTTTACACTTTATGCTTCCGGCTCGTATGTTGTGTGGAATTGTGAGCGGATAACAATTTCACACAGGAAACAGCTATGACCATGATTACGCCAAGCTCTAGCTAGAGGTCGACCAATTCTCATGTTTGACAGCTTATCATCGCAGATCCGGGCAACGTTGTTGCATTGCTGCAGGCGCAGAACTGGTAGGTATGGCAGATCTATACATTGAATCAATATTGGCAATTAGCCATATTAGTCATTGGTTATATAGCATAAATCAATATTGGCTATTGGCCATTGCATACGTTGTATCTATATCATAATATGTACATTTATATTGGCTCATGTCCAATATGACCGCCATGTTGACATTGATTATTGACTAGTTATTAATAGTAATCAATTACGGGGTCATTAGTTCATAGCCCATATATGGAGTTCCGCGTTACATAACTTACGGTAAATGGCCCGCCTGGCTGACCGCCCAACGACCCCCGCCCATTGACGTCAATAATGACGTATGTTCCCATAGTAACGCCAATAGGGACTTTCCATTGACGTCAATGGGTGGAGTATTTACGGTAAACTGCCCACTTGGCAGTACATCAAGTGTATCATATGCCAAGTCCGCCCCCTATTGACGTCAATGACGGTAAATGGCCCGCCTGGCATTATGCCCAGTACATGACCTTACGGGACTTTCCTACTTGGCAGTACATCTACGTATTAGTCATCGCTATTACCATGGTGATGCGGTTTTGGCAGTACACCAATGGGCGTGGATAGCGGTTTGACTCACGGGGATTTCCAAGTCTCCACCCCATTGACGTCAATGGGAGTTTGTTTTGGCACCAAAATCAACGGGACTTTCCAAAATGTCGTAATAACCCCGCCCCGTTGACGCAAATGGGCGGTAGGCGTGTACGGTGGGAGGTCTATATAAGCAGAGCTCGTTTAGTGAACCGTCAGATCCTCACTCTCTTCCGCATCGCTGTCTGCGAGGGCCAGCTGTTGGGCTCGCGGTTGAGGACAAACTCTTCGCGGTCTTTCCAGTACTCTTGGATCGGAAACCCGTCGGCCTCCGAACGGTACTCCGCCACCGAGGGACCTGAGCGAGTCCGCATCGACCGGATCGGAAAACCTCTCGAGAAAGGCGTCTAACCAGTCACAGTCGCAAGGTAGGCTGAGCACCGTGGCGGGCGGCAGCGGGTGGCGGTCGGGGTTGTTTCTGGCGGAGGTGCTGCTGATGATGTAATTAAAGTAGGCGGTSEQ ID NO: 132 ATGCCAAGTGGTCCCAGGCTGAAAATGTGCTCACCCAGTCTCCSEQ ID NO: 133 ATGCCAAGTGGTCCCAGGCTGATATTGTGATGACCCAGGCTGCSEQ ID NO: 134 ATGCCAAGTGGTCCCAGGCTCAAATTGTTCTCACCCAGTCTCCSEQ ID NO: 135 ATGCCAAGTGGTCCCAGGCTAGTATTGTGATGACCCAGACTCCSEQ ID NO: 136 GGGAAGATGAAGACAGATGGTGCAGCCACAGC SEQ ID NO: 137GTAAGCGCTAGCGCCTCAACGAAGGGCCCATCTGTCTTTCCCCTGGCCCC SEQ ID NO: 138GTAAGCGAATTCACAAGATTTGGGCTCAACTTTCTTG SEQ ID NO: 139GCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCAGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAAAGTTGAGCCCAAATCTTGTSEQ ID NO: 140ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSC SEQ ID NO: 141CTTGAGCCGGCGGATGGTCGAGGTGAGGTGTGGCAGGCTTGAGATCCAGCTGTTGGGGTGAGTACTCCCTCTCAAAAGCGGGCATTACTTCTGCGCTAAGATTGTCAGTTTCCAAAAACGAGGAGGATTTGATATTCACCTGGCCCGATCTGGCCATACACTTGAGTGACAATGACATCCACTTTGCCTTTCTCTCCACAGGTGTCCACTCCCAGGTCCAAGTTTGCCGCCACCATGGAGACAGACACACTCCTGCTATGGGTACTGCTGCTCTGGGTTCCAGGTTCCACTGGCGGAGACGGAGCTTACGGGCCCATCGGTCTTCCCCCTGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCTCTGACCAGCGGCGTGCACACCTTCCCAGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAACTTCGGCACCCAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGACAGTTGAGCGCAAATGTTGTGTCGAGTGCCCACCGTGCCCAGCACCACCTGTGGCAGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGACGTGAGCCACGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCACGGGAGGAGCAGTTCAACAGCACGTTCCGTGTGGTCAGCGTCCTCACCGTTGTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGGCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAACCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACACCTCCCATGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCCGGGAAATGATCCCCCGACCTCGACCTCTGGCTAATAAAGGAAATTTATTTTCATTGCAATAGTGTGTTGGAATTTTTTGTGTCTCTCACTCGGAAGGACATATGGGAGGGCAAATCATTTGGTCGAGATCCCTCGGAGATCTCTAGCTAGAGCCCCGCCGCCGGACGAACTAAACCTGACTACGGCATCTCTGCCCCTTCTTCGCGGGGCAGTGCATGTAATCCCTTCAGTTGGTTGGTACAACTTGCCAACTGAACCCTAAACGGGTAGCATATGCTTCCCGGGTAGTAGTATATACTATCCAGACTAACCCTAATTCAATAGCATATGTTACCCAACGGGAAGCATATGCTATCGAATTAGGGTTAGTAAAAGGGTCCTAAGGAACAGCGATGTAGGTGGGCGGGCCAAGATAGGGGCGCGATTGCTGCGATCTGGAGGACAAATTACACACACTTGCGCCTGAGCGCCAAGCACAGGGTTGTTGGTCCTCATATTCACGAGGTCGCTGAGAGCACGGTGGGCTAATGTTGCCATGGGTAGCATATACTACCCAAATATCTGGATAGCATATGCTATCCTAATCTATATCTGGGTAGCATAGGCTATCCTAATCTATATCTGGGTAGCATATGCTATCCTAATCTATATCTGGGTAGTATATGCTATCCTAATTTATATCTGGGTAGCATAGGCTATCCTAATCTATATCTGGGTAGCATATGCTATCCTAATCTATATCTGGGTAGTATATGCTATCCTAATCTGTATCCGGGTAGCATATGCTATCCTAATAGAGATTAGGGTAGTATATGCTATCCTAATTTATATCTGGGTAGCATATACTACCCAAATATCTGGATAGCATATGCTATCCTAATCTATATCTGGGTAGCATATGCTATCCTAATCTATATCTGGGTAGCATAGGCTATCCTAATCTATATCTGGGTAGCATATGCTATCCTAATCTATATCTGGGTAGTATATGCTATCCTAATTTATATCTGGGTAGCATAGGCTATCCTAATCTATATCTGGGTAGCATATGCTATCCTAATCTATATCTGGGTAGTATATGCTATCCTAATCTGTATCCGGGTAGCATATGCTATCCTCACGATGATAAGCTGTCAAACATGAGAATTAATTCTTGAAGACGAAAGGGCCTCGTGATACGCCTATTTTTATAGGTTAATGTCATGATAATAATGGTTTCTTAGACGTCAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGTATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTGTTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGCAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATTTAAAACTTCATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATCCCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCATTGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGTTCTTTCCTGCGTTATCCCCTGATTCTGTGGATAACCGTATTACCGCCTTTGAGTGAGCTGATACCGCTCGCCGCAGCCGAACGACCGAGCGCAGCGAGTCAGTGAGCGAGGAAGCGTACATTTATATTGGCTCATGTCCAATATGACCGCCATGTTGACATTGATTATTGACTAGTTATTAATAGTAATCAATTACGGGGTCATTAGTTCATAGCCCATATATGGAGTTCCGCGTTACATAACTTACGGTAAATGGCCCGCCTGGCTGACCGCCCAACGACCCCCGCCCATTGACGTCAATAATGACGTATGTTCCCATAGTAACGCCAATAGGGACTTTCCATTGACGTCAATGGGTGGAGTATTTACGGTAAACTGCCCACTTGGCAGTACATCAAGTGTATCATATGCCAAGTCCGCCCCCTATTGACGTCAATGACGGTAAATGGCCCGCCTGGCATTATGCCCAGTACATGACCTTACGGGACTTTCCTACTTGGCAGTACATCTACGTATTAGTCATCGCTATTACCATGGTGATGCGGTTTTGGCAGTACACCAATGGGCGTGGATAGCGGTTTGACTCACGGGGATTTCCAAGTCTCCACCCCATTGACGTCAATGGGAGTTTGTTTTGGCACCAAAATCAACGGGACTTTCCAAAATGTCGTAATAACCCCGCCCCGTTGACGCAAATGGGCGGTAGGCGTGTACGGTGGGAGGTCTATATAAGCAGAGCTCGTTTAGTGAACCGTCAGATCCTCACTCTCTTCCGCATCGCTGTCTGCGAGGGCCAGCTGTTGGGCTCGCGGTTGAGGACAAACTCTTCGCGGTCTTTCCAGTACTCTTGGATCGGAAACCCGTCGGCCTCCGAACGGTACTCCGCCACCGAGGGACCTGAGCGAGTCCGCATCGACCGGATCGGAAAACCTCTCGAGAAAGGCGTCTAACCAGTCACAGTCGCAAGGTAGGCTGAGCACCGTGGCGGGCGGCAGCGGGTGGCGGTCGGGGTTGTTTCTGGCGGAGGTGCTGCTGATGATGTAATTAAAGTAGGCGGT SEQ ID NO: 142GGGTTCCAGGTTCCACTGGCGAGGTCCAGCTGCAACAATCTGG SEQ ID NO: 143GGGTTCCAGGTTCCACTGGCCAGGTCCAAGTGCAGCAGCCTGG SEQ ID NO: 144GGGTTCCAGGTTCCACTGGCGAGATCCAGCTGCAGCAGTCTGG SEQ ID NO: 145GGGTTCCAGGTTCCACTGGCGAAGTGAAGCTTGAGGAGTCTGG SEQ ID NO: 146GGGTTCCAGGTTCCACTGGCCAGGTCCAACTGCAGCAGCCTGG SEQ ID NO: 147GGGGCCAGGGGAAAGACAGATGGGCCCTTCGTTGAGGC SEQ ID NO.: 148RSX_(1a)X_(2a)SLLHSNGX_(3a)TYLYX_(1a) is a neutral hydrophilic amino acidX_(3a) is an hydrophobic amino acid or asparagine.X_(2a) is lysine or glutamic acid SEQ ID NO.: 149 X_(1b)MSNLASwherein X_(1b) is a basic amino acid. SEQ ID NO.: 150 RX_(1c)SNLX_(2c)Swherein X_(1c) is methionine or threoninewherein X_(2c) is an hydrophobic amino acid. SEQ ID NO.: 151X_(1d)QX_(2d)LEX_(3d)PX_(4d)Twherein X_(1d) is an hydrophobic amino acid;wherein X_(2d) is abasic amino acid;wherein X_(3d) is tyrosine or leucine, and;wherein X_(4d) is an aromatic amino acid. SEQ ID NO.: 152 QQWSSNPX_(1e)TWherein X_(1e) is proline or leucine. SEQ ID NO.: 153GYTFX_(1f)X_(2f)YX_(3f)MX wherein X_(1f) is threonine or asparagine;wherein X_(2f) is threonine, arginine, serine or aspartic acid;wherein X_(3f) is tryptophan, asparagine, aspartic acid orglutamic acid, and;wherein X_(4f) is tyrosine, histidine or aspartic acid. SEQ ID NO.: 154GYTFTDYX_(5f)MH Wherein X_(5f) is an acidic amino acid. SEQ ID NO.: 155LINPX_(1g)NX_(2g)RX_(3g)NWherein X_(1g) is a neutral hydrophilic amino acid;Wherein X_(2g) is alanine or glycine, and;Wherein X_(3g) is proline ot threonine. SEQ ID NO.: 156 X_(1h)IDPETGGTAWherein X_(1h) is alanine or threonine. SEQ ID NO.: 157EIX_(1i)PX_(2i)X_(3i)SX_(4i)X_(5i)NWherein X_(1i) is aspartic acid or asparagine;Wherein X_(2i) is aspartic acid or serine;Wherein X_(3i) is aspartic acid or serine;Wherein X_(4i) is tyrosine or threonine, and;Wherein X_(5i) is threonine or isoleucine. SEQ ID NO.: 158TX_(1j)FYYX_(2j)X_(3j)X_(4j)NYDVGFAYWherein X_(1j) is a neutral hydrophilic amino acid;Wherein X_(2j) is a neutral hydrophilic amino acid;Wherein X_(3j) is tyrosine or histidine, and;Wherein X_(4j) is tyrosine or serine. SEQ ID NO.: 159GTAAGCGAATTCATGGTGAAAACTAGAAGAGACGC SEQ ID NO.: 160GTAAGCAAGCTTTTAGCCGTGGAAGCGGAACAGGSEQ ID NO.: 161 (25B02 variable light chain DNA)AACATCCAGATGACCCAGTCTCCAGCCTCCCTATCTGCATCTGTGGGAGAAACTGTCACCATCACATGTCGAGCAAGTGAGAATATTTACAGTTATTTAGCATGGTATCAACAGAAGCAGGGAAAATCTCCTCAGCTCCTGGTCTATAATGCAAAAACCTTACCAGAAGGTGTGTCAGTAAGGTTCAGTGGCAGTGGATCAGGCACACAGTTTTCTCTGAAGATCAACAACCTGCAGCCTGAAGATTTTGGGAGTTATCACTGTCAACATCATTATGGTGTTCCTCTTACGTTCGGTTCTGGGACCAAGCTGGAGTTGAAASEQ ID NO.: 162 (25B02 variable light chain amino acids)NIQMTQSPASLSASVGETVTITCRASENIYSYLAWYQQKQGKSPQLLVYNAKTLPEGVSVRFSGSGSGTQFSLKINNLQPEDFGSYHCQHHYGVPLTFGSGTKLELKSEQ ID NO.: 163 (25B02 variable heavy chain DNA)CAGGTGAAGCTTCAGCAGTCCGGGGCTGAGCTGGCAAGACCTGGGGCTTCAGTGAAGTTTTCCTGCAAGGCTTCTGGCTACACCTTCACTAGGAACTGGATACAGTGGGTAAAACAGAGGCCTGGACAGGGTCTGGAATGGATTGGGGCTATTTATCCTGGAAATGGTGATAGTAGGTATACTCAGAAGTTCAAGGGCAAGGCCACATTGACTGCAGATAAATCCTCGAACACAGCCTACATGCAACTCAGCGGTTTGGCATCTGAGGACTCTGCGGTCTATTACTGTGCAAGATTGGCTGGTAACTACGCTTACTACTTTGACTACTGGGGCCAAGGCACCGCTCTCACAGTCTCCTCASEQ ID NO.: 164 (25B02 variable heavy chain amino acids)QVKLQQSGAELARPGASVKFSCKASGYTFTRNWIQWVKQRPGQGLEWIGAIYPGNGDSRYTQKFKGKATLTADKSSNTAYMQLSGLASEDSAVYYCARLAGNYAYYFDYWGQGT ALTVSSSEQ ID NO.: 165 (25D11 variable light chain DNA)GACATCCAGATGACCCAGTCTCCAGCCTCCCTATCTGCATCTGTGGGAGAAACTGTCACCATCACATGTCGAGCAAGTGGGAATATTCACAATTATTTAGCATGGTATCAACAGAAGCAGGGAAAATCTCCTCAGCTCCTGGTCTATAATGCAAAAACCTTACCAGAAGGTGTGTCAGTAAGGTTCAGTGGCAGTGGATCAGGCACACAGTTTTCTCTGAAGATCAACAACCTGCAGCCTGAAGATTTTGGGAGTTATCACTGTCAACATCATTATGGTGTTCCTCTTACGTTCGGTTCTGGGACCAAGCTGGAGTTGAAASEQ ID NO.: 166 (25D11 variable light chain amino acids)DIQMTQSPASLSASVGETVTITCRASGNIHNYLAWYQQKQGKSPQLLVYNAKTLPEGVSVRFSGSGSGTQFSLKINNLQPEDFGSYHCQHHYGVPLTFGSGTKLELKSEQ ID NO.: 167 (25D11 variable heavy chain DNA)CAGGTGAAGCTTCAGCAGTCCGGGGCTGAGCTGGCAAGACCTGGGGCTTCAGTGAAGTTTTCCTGCAAGGCTTCTGGCTACACCTTCACTAGGAACTGGATACAGTGGGTAAAACAGAGGCCTGGACAGGGTCTGGAATGGATTGGGGCTATTTATCCTGGAAATGGTGATAGTAGGTATACTCAGAAGTTCAAGGGCAAGGCCACATTGACTGCAGATAAATCCTCGAACACAGCCTACATGCAACTCAGCGGTTTGGCATCTGAGGACTCTGCGGTCTATTACTGTGCAAGATTGGCTGGTAACTACGCTTACTACTTTGACTACTGGGGCCAAGGCACCGCTCTCACAGTCTCCTCASEQ ID NO.: 168 (25D11 variable heavy chain amino acids)QVKLQQSGAELARPGASVKFSCKASGYTFTRNWIQWVKQRPGQGLEWIGAIYPGNGDSRYTQKFKGKATLTADKSSNTAYMQLSGLASEDSAVYYCARLAGNYAYYFDYWGQGT ALTVSSSEQ ID NO.: 169 (25E10 variable light chain DNA)GACATCCAGATGACCCAGTCTCCAGCCTCCCTATCTGCATCTGTGGGAGAAACTGTCACCATCACATGTCGAGCAAGTGGGAATATTCACAATTATTTAGCATGGTATCAGCAGAAACAGGGAAAATCTCCTCAGCTCCTGGTCTATAATGCAAAAACCCTAGCAGATGGTGTGCCATCAAGGTTCAGTGGCAGTGGATCAGGAACACAATATTCTCTCAAGATCAACAGCCTGCAGCCTGAAGATTTTGGGAGTTATTACTGTCAACATCATTACGGTGCTCCTCTTACGTTCGGTGCTGGGACCAAGGTGGAGCTGAAASEQ ID NO.: 170 (25E10 variable light chain amino acids)DIQMTQSPASLSASVGETVTITCRASGNIHNYLAWYQQKQGKSPQLLVYNAKTLADGVPSRFSGSGSGTQYSLKINSLQPEDFGSYYCQHHYGAPLTFGAGTKVELKSEQ ID NO.: 171 (25E10 variable heavy chain DNA)GATGTGCAGCTGCAACAATCTGGGGCTGAGCTGGCAAGACCTGGGGCTTCAGTGAAGTTTTCCTGCAAGGCTTCTGGCTACACCTTTACTAGGAACTGGATACAGTGGGTTAAACAGAGGCCTGGACAGGGTCTGGAATGGATTGGGGCTGTTTATCCTGGAAATGGTGATAGTAGGTATACTCAGAAGTTCAAGGGCAAGGCCACATTGACTGCAGATAAATCCTCCAGCACAGCCTACATGCAACTCAACAGTTTGTCATCTGAGGACTCTGCGGTCTATTACTGCGCAAGATTGGCTGGTAACTACGCTTACTACTTTGACTACTGGGGCCAAGGCACCGCTCTCACAGTCTCCTCASEQ ID NO.: 172 (25E10 variable heavy chain amino acids)DVQLQQSGAELARPGASVKFSCKASGYTFTRNWIQWVKQRPGQGLEWIGAVYPGNGDSRYTQKFKGKATLTADKSSSTAYMQLNSLSSEDSAVYYCARLAGNYAYYFDYWGQGT ALTVSSSEQ ID NO.: 173 RASENIYSYLA SEQ ID NO.: 174 NAKTLPE SEQ ID NO.: 175QHHYGVPLT SEQ ID NO.: 176 GYTFTRNWIQ SEQ ID NO.: 177 AIYPGNGDSRSEQ ID NO.: 178 ARLAGNYAYYFDY SEQ ID NO.: 179 RASGNIHNYLASEQ ID NO.: 180 NAKTLPE SEQ ID NO.: 181 QHHYGVPLT SEQ ID NO.: 182GYTFTRNWIQ SEQ ID NO.: 183 AIYPGNGDSR SEQ ID NO.: 184 ARLAGNYAYYFDYSEQ ID NO.: 185 RASGNIHNYLA SEQ ID NO.: 186 NAKTLAD SEQ ID NO.: 187QHHYGAPLT SEQ ID NO.: 188 GYTFTRNWIQ SEQ ID NO.: 189 AVYPGNGDSRSEQ ID NO.: 190 ARLAGNYAYYFDY

TABLE 5A Anti-siglec-15 heavy chain variable sequences. ID FR1 CDR-H1FR2 CDR-H2 25E6 QVQLQQPGAELAKPGASVKLSCKAS GYTFNTYNMY WLKQRPGQGLEWIGGIDPSNGDTK 25H10 QVQLQQPGAELAKPGASVKLSCKAS GYTFNTYNMY WLKQRPGQGLEWIGGIDPSNGDTK 25H11 QVQLQQPGAELAKPGASVKLSCKAS GYTFNTYNMY WLKQRPGQGLEWIGGIDPSNGDTK 25A3 QVQLQQSRAELVKPGASVKLSCKAS GYTFTSYWMH WVKQRPGQGLEWIGLINPTNGRTN 25A5 QVQVQQPGAELVKPGASVKLSCKAS GYTFTSYWMH WVKQRPGQGLEWIGLINPSNARTN 25A11 QVQVQQPGAEIVRPGASVKLSCKAS GYTFTSYWMH WVKQRPGQGLEWIGLINPTNGRTN 25B4 QVQVQQPGAEIVRPGASVKLSCKAS GYTFTSYWMH WVKQRPGQGLEWIGLINPTNGRTN 25B12 QVQLQQSRAELVKPGASVKLSCKAS GYTFTSYWMH WVKQRPGQGLEWIGLINPTNGRTN 25C9 QVQVQQPGAELVKPGASVKLSCKAS GYTFTSYWMH WVKQRPGQGLEWIGLINPSNARTN 25C10 QVQVQQPGAELVKPGASVKLSCKAS GYTFTSYWMH WVKQRPGQGLEWIGLINPSNARTN 25D3 QVQLQQSRAELVKPGASVKLSCKAS GYTFTSYWMH WVKQRPGQGLEWIGLINPTNGRTN 25D4 QVQVQQPGAEIVRPGASVKLSCKAS GYTFTSYWMH WVKQRPGQGLEWIGLINPTNGRTN 25D5 QVQVQQPGAEIVRPGASVKLSCKAS GYTFTSYWMH WVKQRPGQGLEWIGLINPTNGRTN 25D6 QVQVQQPGAEIVRPGASVKLSCKAS GYTFTSYWMH WVKQRPGQGLEWIGLINPTNGRTN 25D8 QVQVQQPGAELVKPGASVKLSCKAS GYTFTSYWMH WVKQRPGQGLEWIGLINPSNARTN 25D10 QVQVQQPGAEIVRPGASVKLSCKAS GYTFTSYWMH WVKQRPGQGLEWIGLINPTNGRTN 25E7 QVQVQQPGTELVKPGASVKLSCKAS GYTFTTYWMH WVKQRPGQGLEWIGLINPTNGRTN 25E8 QVQLQQSRAELVKPGASVKLSCKAS GYTFTSYWMH WVKQRPGQGLEWIGLINPTNGRTN 25E12 QVQVQQPGAEIVRPGASVKLSCKAS GYTFTSYWMH WVKQRPGQGLEWIGLINPTNGRTN 25F2 QVQVQQPGAEIVRPGASVKLSCKAS GYTFTSYWMH WVKQRPGQGLEWIGLINPTNGRTN 25F3 QVQVQQPGAELVKPGASVKLSCKAS GYTFTSYWMH WVKQRPGQGLEWIGLINPSNARTN 25F5 QVQVQQPGTELVKPGASVKLSCKAS GYTFTTYWMH WVKQRPGQGLEWIGLINPTNGRTN 25F6 QVQVQQPGAELVKPGASVKLSCKAS GYTFTSYWMH WVKQRPGQGLEWIGLINPSNARTN 25F7 EIQLQQSGTELVKPGASVKLSCKAS GYTFTTYWMH WVKQRPGQGLEWIGLINPTNGRTN 25F9 QVQVQQPGAEIVRPGASVKLSCKAS GYTFTSYWMH WVKQRPGQGLEWIGLINPTNGRTN 25F10 QVQVQQPGTELVKPGASVKLSCKAS GYTFTTYWMH WVKQRPGQGLEWIGLINPTNGRTN 25F11 QVQVQQPGAEIVRPGASVKLSCKAS GYTFTSYWMH WVKQRPGQGLEWIGLINPTNGRTN 25F12 QVQVQQPGAEIVRPGASVKLSCKAS GYTFTSYWMH WVKQRPGQGLEWIGLINPTNGRTN 25G3 QVQVQQPGAEIVRPGASVKLSCKAS GYTFTSYWMH WVKQRPGQGLEWIGLINPTNGRTN 25G4 QVQVQQPGAEIVRPGASVKLSCKAS GYTFTSYWMH WVKQRPGQGLEWIGLINPTNGRTN 25G7 QVQVQQPGAEIVRPGASVKLSCKAS GYTFTSYWMH WVKQRPGQGLEWIGLINPTNGRTN 25GB QVQVQQPGAEIVRPGASVKLSCKAS GYTFTSYWMH WVKQRPGQGLEWIGLINPTNGRTN 25H1 QVQVQQPGTELVKPGASVKLSCKAS GYTFTTYWMH WVKQRPGQGLEWIGLINPTNGRTN 25H2 QVQVQQPGAEIVRPGASVKLSCKAS GYTFTSYWMH WVKQRPGQGLEWIGLINPTNGRTN 25H5 QVQLQQSRAELVKPGASVKLSCKAS GYTFTSYWMH WVKQRPGQGLEWIGLINPTNGRTN 25H6 QVQLQQSGAELVKPGASVKLSCKAS GYTFTSYWMH WVKQRPGQGLEWIGLINPSNGRPN 25H7 QVQVQQPGAELVKPGASVKLSCKAS GYTFTSYWMH WVKQRPGQGLEWIGLINPSNARTN 25H8 QVQVQQPGAEIVRPGASVKLSCKAS GYTFTSYWMH WVKQRPGQGLEWIGLINPTNGRTN 25B2 QVKLQQSGAELARPGASVKFSCKAS GYTFTRNWIQ WVKQRPGQGLEWIGAIYPGNGDSR 25D11 QVKLQQSGAELARPGASVKFSCKAS GYTFTRNWIQ WVKQRPGQGLEWIGAIYPGNGDSR 25E10 DVQLQQSGAELARPGASVKFSCKAS GYTFTRNWIQ WVKQRPGQGLEWIGAVYPGNGDSR 25E5 EVKLEESGGGLVQPGGSLKLSCAAS GFDFSKDWMS WVRQAPGKGLEWIGEINPDSSTIN 25B6 QAYLQQSGVELVRPGASVTLSCKAS GYTFTDYDMH WVKQTPVHGLEWIGTIDPETGGTA 25B11 EIQLQQSGVELVRPGASVTLSCKAS GYTFTDYDMH WVKQTPVHGLEWIGTIDPETGGTA 25E9 EIQLQQSGVELVRPGASVTLSCKAS GYTFTDYDMH WVKQTPVHGLEWIGTIDPETGGTA 25C1 EIQLQQSGAELVRPGASVTLSCKAS GYTFTDYEMH WVKQTPVHGLEWIGAIDPETGGTA 25B8 EIQLQQSGVELVRPGASVTLSCKAS GYTFTDYDMH WVKQTPVHGLEWIGTIDPETGGTA 25A1 EVQLQQSGTELVRPGSSVKISCKAS GYTFTRYWMD WVKQRPGQGLEWIGEIDPSDSYTN ID FR3 CDR-H3 FR4 25E6 INEKFKNKATLTVDKSSSTAYMQLSGLTSEDSAVYYCTSH------------TY WGQGTLVTVSA 25H10INEKFKNKATLTVDKSSSTAYMQLSGLTSEDSAVYYC TSH------------TY WGQGTLVTVSA25H11 INEKFKNKATLTVDKSSSTAYMQLSGLTSEDSAVYYC TSH------------TYWGQGTLVTVSA 25A3 YNEKFKNKATLTVDKSSSTAYMQLSSLTSEDSAVYYC ARGGDGDYF------DYWGQGTTLTVSS 25A5 YNEKFNTKATLTVDKSSSTAYMQLSSLTSEDSAVYYC ARGGDGDYF------DYWGQGTTLTVSS 25A11 YNEKFKSKATLTVDKSSSTAYMQLSSLTSEDSAVYYCARGGDGDYF------DY WGQGTTLTVSS 25B4 YNEKFKSKATLTVDKSSSTAYMQLSSLTSEDSAVYYCARGGDGDYF------DY WGQGTTLTVSS 25B12YNEKFKNKATLTVDKSSSTAYMQLSSLTSEDSAVYYC ARGGDGDYF------DY WGQGTTLTVSS 25C9YNEKFNTKATLTVDKSSSTAYMQLSSLTSEDSAVYYC ARGGDGDYF------DY WGQGTTLTVSS25C10 YNEKFNTKATLTVDKSSSTAYMQLSSLTSEDSAVYYC ARGGDGDYF------DYWGQGTTLTVSS 25D3 YNEKFKNKATLTVDKSSSTAYMQLSSLTSEDSAVYYC ARGGDGDYF------DYWGQGTTLTVSS 25D4 YNEKFKSKATLTVDKSSSTAYMQLSSLTSEDSAVYYC ARGGDGDYF------DYWGQGTTLTVSS 25D5 YNEKEKSKATLTVDKSSSTAYMQLSSLTSEDSAVYYC ARGGDGDYF------DYWGQGTTLTVSS 25D6 YNEKEKSKATLTVDKSSSTAYMQLSSLTSEDSAVYYC ARGGDGDYF------DYWGQGTTLTVSS 25D8 YNEKENTKATLTVDKSSSTAYMQLSSLTSEDSAVYYC ARGGDGDYF------DYWGQGTTLTVSS 25D10 YNEKEKSKATLTVDKSSSTAYMQLSSLTSEDSAVYYCARGGDGDYF------DY WGQGTTLTVSS 25E7 YNEKFKSKATLTVDKSSSTAYMQLSSLTSEDSAVYYCARGGDGDYF------DY WGQGTTLTVSS 25E8 YNEKEKNKATLTVDKSSSTAYMQLSSLTSEDSAVYYCARGGDGDYF------DY WGQGTTLTVSS 25E12YNEKFKSKATLTVDKSSSTAYMHLSSLTSEDSAVYYC ARGGDGDYF------DY WGQGTTLTVSS 25F2YNEKFKSKATLTVDKSSSTAYMQLSSLTSEDSAVYYC ARGGDGDYF------DY WGQGTTLTVSS 25F3YNEKFNTKATLTVDKSSSTAYMQLSSLTSEDSAVYYC ARGGDGDYF------DY WGQGTTLTVSS 25F5YNEKFKSKATLTVDKSSSTAYMQLSSLTSEDSAVYYC ARGGDGDYF------DY WGQGTTLTVSS 25F6YNEKFNTKATLTVDKSSSTAYMQLSSLTSEDSAVYYC ARGGDGDYF------DY WGQGTTLTVSS 25F7YNEKFKSKATLTVDKSSSTAYMQLSSLTSEDSAVYYC ARGGDGDYF------DY WGQGTTLTVSS 25F9YNEKFKSKATLTVDKSSSTAYMQLSSLTSEDSAVYYC ARGGDGDYF------DY WGQGTTLTVSS25E10 YNEKEKSKATLTVDKSSSTAYMQLSSLTSEDSAVYYC ARGGDGDYF------DYWGQGTTLTVSS 25E11 YNEKFKSKATLTVDKSSSTAYMQLSSLTSEDSAVYYCARGGDGDYF------DY WGQGTTLTVSS 25E12YNEKFKNKATLTVDKSSSTAYMQLSSLTSEDSAVYYC ARGGDGDYF------DY WGQGTTLTVSS 25G3YNEKFKSKATLTVDKSSSTAYMQLSSLTSEDSAVYYC ARGGDGDYF------DY WGQGTTLTVSS 25G4YNEKFKSKATLTVDKSSSTAYMQLSSLTSEDSAVYYC ARGGDGDYF------DY WGQGTTLTVSS 25G7YNEKFKSKATLTVDKSSSTAYMQLSSLTSEDSAVYYC ARGGDGDYF------DY WGQGTTLTVSS 25GBYNEKFKSKATLTVDKSSSTAYMQLSSLTSEDSAVYYC ARGGDGDYF------DY WGQGTTLTVSS 25H1YNEKFKSKATLTVDKSSSTAYMQLSSLTSEDSAVYYC ARGGDGDYF------DY WGQGTTLTVSS 25H2YNEKFKSKATLTVDKSSSTAYMQLSSLTSEDSAVYYC ARGGDGDYF------DY WGQGTTLTVSS 25H5YNEKFKNKATLTVDKSSSTAYMQLSSLTSEDSAVYYC ARGGDGDYF------DY WGQGTTLTVSS 25H6YNERFKTKATLTVDKSSSTAYMHLSSLTSEDSAVYYC ARGGDGDYF------DY WGQGTTLTVSS 25H7YNEKFNTKATLTVDKSSSTAYMQLSSLTSEDSAVYYC ARGGDGDYF------DY WGQGTTLTVSS 25H8YNEKFKSKATLTVDKSSSTAYMQLSSLTSEDSAVYYC ARGGDGDYF------DY WGQGTTLTVSS 25B2YTQKFKGKATLTADKSSNTAYMQLSGLASEDSAVYYC ARLAGNYAYYF----DY WGQGTALTVSS25D11 YTQKFKGKATLTADKSSNTAYMQLSGLASEDSAVYYC ARLAGNYAYYF----DYWGQGTALTVSS 25E10 YTQKFKGKATLTADKSSSTAYMQLNSLSSEDSAVYYCARLAGNYAYYF----DY WGQGTALTVSS 25E5 YAPSLKDKFIISRENAKNTLYLQMSKVRSEDTALYYCSRLEDYEDWYF----DV WGAGTTVTVSS 25B6 YNQKFKGKATLTADRSSTTAYMELSSLTSEDSAVYYCTSFYYTYSNYDVGF-AY WGQGTLVTVSA 25B11YNQKFKGKATLTADRSSTTAYMELSSLTSEDSAVYYC TSFYYTYSNYDVGF-AY WGQGTLVTVSA 25E9YNQKFKGKATLTADRSSTTAYMELSSLTSEDSAVYYC TSFYYTYSNYDVGF-AY WGQGTLVTVSA 25C1YNQKFKGKATLTADKSSSTAYMELSSLTSEDSAVYYC TSFYYTYYNYDVGF-AY WGQGTLVTVSA 25B8YNQKFKGKATLTADRSSTTAYMELSSLTSEDSAVYYC TTFYYSHYNYDVGF-AY WGQGTLVTVSA 25A1YNQKFKGKATLTVDKFSRTAYMELSSLTSEDSAVYYC ARSGAYSSDYSYDGFAY WGQGTLVTVSA

TABLE 5B Anti-siglec-15 light chain variable sequences. ID FR1 CDR-L1FR2 CDR-L2 25E6 SIVMTQTPKFLLVSAGDRVTITC KASQSVS-----NAVA WYQQKPGQSPKLLIYYTSNRYT 25H10 SIVMTQTPKFLLVSAGDRVTITC KASQSVS-----NAVA WYQQKPGQSPKLLIYYTSNRYT 25H11 SIVMTQTPKFLLVSAGDRVTITC KASQSVS-----NAVA WYQQKPGQSPKLLIYYTSNRYT 25A3 DIVMTQAAFSNPVTLGTSASISC RSSKSLLHSNGITYLY WYLQKPGQSPQLLIYQMSNLAS 25A5 DIVMTQAAFSNPVTLGTSASISC RSSKSLLHSNGITYLY WYLQKPGQSPQLLIYQMSNLAS 25A11 DIVMTQAAFSNPVTLGTSASISC RSSKSLLHSNGITYLY WYLQKPGQSPQLLIYQMSNLAS 25B4 DIVMTQAAFSNPVTLGTSASISC RSSKSLLHSNGITYLY WYLQKPGQSPQLLIYQMSNLAS 25B12 DIVMTQAAFSNPVTLGTSASISC RSSKSLLHSNGITYLY WYLQKPGQSPQLLIYQMSNLAS 25C9 DIVMTQAAFSNPVTLGTSASISC RSSKSLLHSNGITYLY WYLQKPGQSPQLLIYQMSNLAS 25C10 DIVMTQAAFSNPVTLGTSASISC RSSKSLLHSNGITYLY WYLQKPGQSPQLLIYQMSNLAS 25D3 DIVMTQAAFSNPVTLGTSASISC RSSKSLLHSNGITYLY WYLQKPGQSPQLLIYQMSNLAS 25D4 DIVMTQAAFSNPVTLGTSASISC RSSKSLLHSNGITYLY WYLQKPGQSPQLLIYQMSNLAS 25D5 DIVMTQAAFSNPVTLGTSASISC RSSKSLLHSNGITYLY WYLQKPGQSPQLLIYQMSNLAS 25D6 DIVMTQAAFSNPVTLGTSASISC RSSKSLLHSNGITYLY WYLQKPGQSPQLLIYQMSNLAS 25D8 DIVMTQAAFSNPVTLGTSASISC RSSKSLLHSNGITYLY WYLQKPGQSPQLLIYQMSNLAS 25D10 DIVMTQAAFSNPVTLGTSASISC RSSKSLLHSNGITYLY WYLQKPGQSPQLLIYQMSNLAS 25E7 DIVMTQAVFSNPVILGTPASISC RSSKSLLHSNGVTYLY WYLQKPGQSPQLLIYQMSNLAS 25E8 DIVMTQAAFSNPVTLGTSASISC RSSKSLLHSNGITYLY WYLQKPGQSPQLLIYQMSNLAS 25E12 DIVMTQAAFSNPVTLGTSASISC RSSESLLHSNGITYLY WYLQKPGQSPQLLIYQMSNLAS 25F2 DIVMTQAAFSNPVTLGTSASISC RSSKSLLHSNGITYLY WYLQKPGQSPQLLIYQMSNLAS 25F3 DIVMTHAAFSNPVTLGTSASISC RSSKSLLHSNGITYLY WYLQKPGQSPQLLIYQMSNLAS 25F5 DIVMTQAVFSNPVILGTPASISC RSSKSLLHSNGVTYLY WYLQKPGQSPQLLIYQMSNLAS 25F6 DIVMTQAAFSNPVTLGTSASISC RSSKSLLHSNGITYLY WYLQKPGQSPQLLIYQMSNLAS 25F7 DIVMTQAAFSNPVTLGTSASISC RSSKSLLHSNGITYLY WYLQKPGQSPQLLIYQMSNLAS 25F9 DIVMTQAAFSNPVTLGTSASISC RSSKSLLHSNGITYLY WYLQKPGQSPQLLIYQMSNLAS 25F10 DIVMTQAVFSNPVILGTPASISC RSSKSLLHSNGVTYLY WYLQKPGQSPQLLIYQMSNLAS 25F11 DIVMTQAAFSNPVTLGTSASISC RSSKSLLHSNGITYLY WYLQKPGQSPQLLIYQMSNLAS 25F12 DIVMTQAAFSNPVTLGTSASISC RSSKSLLHSNGITYLY WYLQKPGQSPQLLIYQMSNLAS 25G3 DIVMTQAAFSNPVTLGTSASISC RSSKSLLHSNGITYLY WYLQKPGQSPQLLIYQMSNLAS 25G4 DIVMTQAAFSNPVTLGTSASISC RSSKSLLHSNGITYLY WYLQKPGQSPQLLIYQMSNLAS 25G7 DIVMTQAAFSNPVTLGTSASISC RSSKSLLHSNGITYLY WYLQKPGQSPQLLIYQMSNLAS 25G8 DIVMTQAAFSNPVTLGTSASISC RSSKSLLHSNGITYLY WYLQKPGQSPQLLIYQMSNLAS 25H1 DIVMTQAAFSNPVTLGTSASISC RSSKSLLHSNGITYLY WYLQKPGQSPQLLIYQMSNLAS 25H2 DIVMTQAAFSNPVTLGTSASISC RSSKSLLHSNGITYLY WYLQKPGQSPQLLIYQMSNLAS 25H5 DIVMTQAAFSNPVTLGTSASISC RSSKSLLHSNGITYLY WYLQKPGQSPQLLIYQMSNLAS 25H6 DIVMTQAAFSNPVTLGTSASISC RSSKSLLHSNGITYLY WYLQKPGQSPQLLIYQMSNLAS 25H7 DIVMTQAAFSNPVTLGTSASISC RSSKSLLHSNGITYLY WYLQKPGQSPQLLIYQMSNLAS 25H8 DIVMTQAAFSNPVTLGTSASISC RSSKSLLHSNGITYLY WYLQKPGQSPQLLIYQMSNLAS 25B2 NIQMTQSPASLSASVGETVTITC RASENIY-----SYLA WYQQKQGKSPQLLVYNAKTLPE 25D11 DIQMTQSPASLSASVGETVTITC RASGNIH-----NYLA WYQQKQGKSPQLLVYNAKTLPE 25E10 DIQMTQSPASLSASVGETVTITC RASGNIH-----NYLA WYQQKQGKSPQLLVYNAKTLAD 25E5 QIVLTQSPTLMSASPGEKVTMTC SASSSV------SYMY WYQQKPRSSPKPWIYRTSNLVS 25B6 DIVMTQAAPSVPVTPGESVSISC RSSKSLLHSNGNTYLY WFLQRPGQSPQLLIYRMSNLAS 25B11 DIVMTQAAPSVPVTPGESVSISC RSTKSLLHSNGNTYLY WFLQRPGQSPQLLIYRMSNLAS 25E9 DIVMTQAAPSVPVTPGESVSISC RSTKSLLHSNGNTYLY WFLQRPGQSPQLLIYRMSNLAS 25C1 DIVMTQAAPSVPVTPGESVSISC RSSKSLLHSNGNTYLY WFLQRPGQSPQLLIYRMSNLAS 25B8 DIVMTQAAPSVPVTPGESVSISC RSTKSLLHSNGNTYLY WFLQRPGQSPQLLIYRMSNLAS 25A1 ENVLTQSPAIMSASPGEKVTISC SASSSV------SYMY WYQQKPGSSPKPWIYRTSNLAS ID FR3 CDR-L3 FR4 25E6 GVPDRFTGSGYGTDFTFTITTVQAEDLAVYFCQQDYTSPWT FGGGTKLEIK 25H10 GVPDRFTGSGYGTDFTFTITTVQAEDLAVYFC QQDYTSPWTFGGGTKLEIK 25H11 GVPDRFTGSGYGTDFTFTITTVQAEDLAVYFC QQDYTSPWT FGGGTKLEIK25A3 GVPDRFSSSGSGTDFTLRISRVEAEDVGVYYC AQNLELPYT FGGGTKLEIK 25A5GVPDRFSSSGSGTDFTLRISRVEAEDVGVYYC AQNLELPYT FGGGTKLEIK 25A11GVPDRFSGSGSGTAFTLRISRVEAEDVGVYYC MQHLEYPYT FGGGTKLEIK 25B4GVPDRFSGSGSGTAFTLRISRVEAEDVGVYYC MQHLEYPYT FGGGTKLEIK 25B12GVPDRFSSSGSGTDFTLRISRVEAEDVGVYYC AQNLELPYT FGGGTKLEIK 25C9GVPDRFSSSGSGTDFTLRISRVEAEDVGVYYC AQNLELPYT FGGGTKLEIK 25C10GVPDRFSSSGSGTDFTLRISRVEAEDVGVYYC AQNLELPYT FGGGTKLEIK 25D3GVPDRFSSSGSGTDFTLRISRVEAEDVGVYYC AQNLELPYT FGGGTKLEIK 25D4GVPDRFSGSGSGTAFTLRISRVEAEDVGVYYC MQHLEYPYT FGGGTKLEIK 25D5GVPDRFSGSGSGTAFTLRISRVEAEDVGVYYC MQHLEYPYT FGGGTKLEIK 25D6GVPDRFSGSGSGTAFTLRISRVEAEDVGVYYC MQHLEYPYT FGGGTKLEIK 25D8GVPDRFSSSGSGTDFTLRISRVEAEDVGVYYC AQNLELPYT FGGGTKLEIK 25D10GVPDRFSGSGSGTAFTLRISRVEAEDVGVYYC MQHLEYPYT FGGGTKLEIK 25E7GVPDRFSSSGSGTDFTLRISRVEAEDVGVYYC MQHLEYPYT FGGGTKLEIK 25E8GVPDRFSSSGSGTDFTLRISRVEAEDVGVYYC AQNLELPYT FGGGTKLEIK 25E12GVPDRFSGSGSGTAFTLRISRVEAEDVGVYYC MQHLEYPYT FGGGTKLEIK 25F2GVPDRFSGSGSGTAFTLRISRVEAEDVGVYYC MQHLEYPYT FGGGTKLEIK 25F3GVPDRFSSSGSGTDFTLRISRVEAEDVGVYYC AQNLELPYT FGGGTKLEIK 25F5GVPDRFSSSGSGTDFTLRISRVEAEDVGVYYC MQHLEYPYT FGGGTKLEIK 25F6GVPDRFSGSGSGTAFTLRISRVEAEDVGVYYC MQHLEYPYT FGGGTKLEIK 25F7GVPDRFSSSGSGTAFTLRISRVEAEDVGVYYC MQHLEYPYT FGGGTKLEIK 25F9GVPDRFSGSGSGTAFTLRISRVEAEDVGVYYC MQHLEYPYT FGGGTKLEIK 25F10GVPDRFSSSGSGTDFTLRISRVEAEDVGVYYC MQHLEYPYT FGGGTKLEIK 25F11GVPDRFSGSGSGTAFTLRISRVEAEDVGVYYC MQHLEYPYT FGGGTKLEIK 25F12GVPDRFSGSGSGTAFTLRISRVEAEDVGVYYC MQHLEYPYT FGGGTKLEIK 25G3GVPDRFSGSGSGTAFTLRISRVEAEDVGVYYC MQHLEYPYT FGGGTKLEIK 25G4GVPDRFSGSGSGTAFTLRISRVEAEDVGVYYC MQHLEYPYT FGGGTKLEIK 25G7GVPDRFSGSGSGTAFTLRISRVEAEDVGVYYC MQHLEYPYT FGGGTKLEIK 25G8GVPDRFSGSGSGTAFTLRISRVEAEDVGVYYC MQHLEYPYT FGGGTKLEIK 25H1GVPDRFSSSGSGTDFTLRISRVEAEDVGVYYC AQNLELPYT FGGGTKLEIK 25H2GVPDRFSGSGSGTAFTLRISRVEAEDVGVYYC MQHLEYPYT FGGGTKLEIK 25H5GVPDRFSSSGSGTDFTLRISRVEAEDVGVYYC AQNLELPYT FGGGTKLEIK 25H6GVPDRFSSSGSGTDFTLRISRVEAEDVGVYYC AQNLEYPYT FGSGTKLEIK 25H7GVPDRFSSSGSGTDFTLRISRVEAEDVGVYYC AQNLELPYT FGGGTKLEIK 25H8GVPDRFSGSGSGTAFTLRISRVEAEDVGVYYC MQHLEYPYT FGGGTKLEIK 25B2GVSVRFSGSGSGTQFSLKINNLQPEDFGSYHC QHHYGVPLT FGSGTKLELK 25D11GVSVRFSGSGSGTQFSLKINNLQPEDFGSYHC QHHYGVPLT FGSGTKLELK 25E10GVPSRFSGSGSGTQYSLKINSLQPEDFGSYYC QHHYGAPLT FGAGTKVELK 25E5GVPVRFSGSGSGTSYSLTISSMEAEDAATYYC QQWSSNPPT FGAGTKLELK 25B6GVPDRFSGSGSGTAFTLRLSRVEAEDVGVYYC MQHLEYPFT FGGGTKLEIK 25B11GVPDRFSGSGSGTAFTLRISRVEAEDVGVYYC MQHLEYPFT FGGGTKLEIK 25E9GVPDRFSGSGSGTAFTLRISRVEAEDVGVYYC MQHLEYPFT FGGGTKLEIK 25C1GVPDRFSGSGSGTAFTLRISRVEAEDVGVYYC MQHLEYPFT FGGGTKLEIK 25B8GVPDRFSGSGSGTAFTLRISRVEAEDVGVYYC MQHLEYPFT FGGGTKLEIK 25A1GVPARFSGSGSGTSYSLTISSMEAEDAATYYC QQWSSNPLT FGAGTKLELK

1-53. (canceled)
 54. A nucleic acid encoding a light chain variabledomain and/or heavy chain variable domain of an antibody thatspecifically binds to Siglec-15 (SEQ ID NO.:2), said light chainvariable domain comprising complementary determining regions having theamino acid sequence set forth in SEQ ID NO.:111, SEQ ID NO.:112 and SEQID NO.:113 and said heavy chain variable domain comprisingcomplementarity determining regions having the amino acid sequence setforth in SEQ ID NO.:114, SEQ ID NO.:115 and SEQ ID NO.:
 116. 55. Avector or vectors comprising the nucleic acid of claim
 54. 56. Thevector or vectors of claim 55, wherein said vector or vectors comprisesa constitutive or an inducible promoter.
 57. An isolated cell comprisingthe nucleic acid of claim
 54. 58. (canceled)
 59. The isolated cell ofclaim 57, wherein said cell is capable of expressing, assembling and/orsecreting an antibody or antigen binding fragment thereof. 60-92.(canceled)
 93. A method of making an antibody comprising culturing thecell of claim 57 so that the antibody is produced.
 94. The method ofclaim 93, comprising recovering the antibody.
 95. The method of claim94, further comprising conjugating the antibody with a therapeutic orcytotoxic agent.
 96. A nucleic acid encoding a light chain variabledomain and/or a heavy chain variable domain of an antibody thatspecifically binds to Siglec-15 (SEQ ID NO.:2), said light chainvariable domain comprising complementary determining regions having theamino acid sequence set forth in SEQ ID NO.:111, SEQ ID NO.:112 and SEQID NO.:113 and said heavy chain variable domain comprisingcomplementarity determining regions having the amino acid sequence setforth in SEQ ID NO.:114, SEQ ID NO.:115 and SEQ ID NO.: 116, wherein atleast one of the light chain variable region and heavy chain variableregion comprises framework amino acids of a human antibody.
 97. A vectoror vectors comprising the nucleic acid of claim
 96. 98. An isolated cellcomprising the vector or vectors of claim
 97. 99. A method of making anantibody comprising culturing the cell of claim 98 so that the antibodyis produced.
 100. The method of claim 99, comprising recovering theantibody.
 101. The method of claim 100, further comprising conjugatingthe antibody with a therapeutic or cytotoxic agent.
 102. A cellconstitutively- or inducibly-expressing an antibody that specificallybinds to Siglec-15, wherein said cell comprises a nucleic acid ornucleic acids encoding a light chain variable domain comprisingcomplementary determining regions having the amino acid sequence setforth in SEQ ID NO.:111, SEQ ID NO.:112 and SEQ ID NO.:113 and a heavychain variable domain comprising complementarity determining regionshaving the amino acid sequence set forth in SEQ ID NO.:114, SEQ IDNO.:115 and SEQ ID NO.:
 116. 103. A method of making an antibodycomprising culturing the cell of claim 102 so that the antibody isproduced.
 104. The method of claim 103, comprising recovering theantibody.
 105. The method of claim 104, further comprising conjugatingthe antibody with a therapeutic or cytotoxic agent.